CN1688206A

CN1688206A - Use of astaxanthin-containing plants or parts of plants of the genus tagetes as animal feed

Info

Publication number: CN1688206A
Application number: CNA038242052A
Authority: CN
Inventors: R·弗拉赫曼; M·绍尔; C·R·朔普费尔; M·克勒布萨特尔; A-M·普法伊费尔; T·卢克; D·弗斯特
Original assignee: BASF SE; BASF Plant Science GmbH; SunGene GmbH
Current assignee: BASF SE; BASF Plant Science GmbH; SunGene GmbH
Priority date: 2002-08-20
Filing date: 2003-08-18
Publication date: 2005-10-26
Anticipated expiration: 2023-08-18
Also published as: CN100364438C; DE10238978A1

Abstract

The invention relates to the use of astaxanthin-containing plants or parts of plants of the genus Tagetes or astaxanthin-containing extracts of astaxanthin-containing plants or parts of plants of the genus Tagetes for oral administration to animals. The invention also relates to methods for producing animal feed preparations, to said animal feed preparations themselves, to a method for the pigmentation of animals or animal products, and to a method for producing pigmented animals and animal products.

Description

The plants of tagetes species of astaxanthin-containing or plant part are as the purposes of feed

The astaxanthin-containing extract that the present invention relates to the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or its part or astaxanthin-containing or its part is used to be administered orally in the purposes of animal, produce the method for animal feed goods, animal feed goods itself, make animal or animal product method of colouring, and relate to the method that produces painted animal or animal product.

Because its painted characteristic, astaxanthin is as the pigment in the animal nutrition, in particular for trout, salmon and shrimp culture.

Mainly produce astaxanthin at present by chemical synthesis process.Pass through to cultivate for example haematococcus pluvialis (Haematococcus pluvialis) of algae at present, perhaps also separate producing a small amount of natural astaxanthin subsequently with biological technique method by the hereditary microorganism of upward optimizing of fermenting.

By the synthesizing astaxanthin that separate to produce or natural astaxanthin is the chemistry that produces by the special preparation technology that is used for prolonging storage life or physically stable, and uses scope and bioavilability is produced the purposes that is used for separately according to expection.

WO 9201754 has described a kind of wild-type plant species summer adonis (Adonis aestivalis) of astaxanthin-containing.In addition, the document also discloses that the summer adonis petal of astaxanthin-containing and its extract as fish food, or as the purposes that is used to make the painted additive of fish in the fish food.

Yet, summer adonis is as being used to make the plant origin of the painted astaxanthin of fish to have such shortcoming in the prior art, be the output of astaxanthin-containing biomass the and therefore output of per unit cultivation region astaxanthin-containing vegetable material is very low, and therefore can only cultivate the astaxanthin-containing vegetable material of acquisition acceptable amount by expensive large tracts of land.This causes expensive in producing of corresponding pigment.

Therefore providing the colouring agent of the shortcoming that no longer has prior art is purpose of the present invention.

The plants of tagetes species of the plants of tagetes species that has been found that astaxanthin-containing as previously mentioned or its part or astaxanthin-containing or the astaxanthin-containing extract of its part can be used in and be administered orally in animal.

In preferred embodiments, the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or its part or astaxanthin-containing or its part is used to make animal painted and make corresponding animal product painted.

Think that preferably the plants of tagetes species of astaxanthin-containing is meant the plants of tagetes species that contains a certain amount of astaxanthin at least in part of plant.Astaxanthin can be with free form, exist with aliphatic acid diester or monoester form.Preferred plants of tagetes species is selected to sow: marigold (Tageteserecta), maidenhair (Tagetes patula) (these two kinds of plants of tagetes species are also referred to as pot marigold), spiceleaf marigold (Tagetes lucida), general woods marigold (Tagetes pringlei), Tagetespalmeri, little marigold (Tagetes minuta), lemon marigold (Tagetes lemmonii), Tagetes signata (Tagetes tenuifolia) or bell marigold (Tagetes campanulata) particularly preferably are marigold or maidenhair.

Think that preferably the part of plants of tagetes species of astaxanthin-containing is meant the plant part that contains a certain amount of astaxanthin at least in a part of this plant part.For example, preferred plant part is flower, capitulum (flower heads) or is floral leaf particularly preferably, is also referred to as petal.

Tagetes wild-type plant astaxanthin-containing not in spending, but contain carotenoid, as lutein and luteole.Yet, have been found that the ability that can give plants of tagetes species generation astaxanthin according to the present invention, for example pass through genetic modification.

In preferred embodiments, for example by causing that in compare with wild type the ketolase activity gives the ability that plants of tagetes species produces astaxanthin by the plants of tagetes species of genetic modification.

Think that the ketolase activity is meant the enzymatic activity of ketolase.

Think that ketolase is meant the protein that has in the enzymatic activity of the optional β that replaces of carotenoid-ionone ring place introducing ketone group.

Particularly, think that ketolase is meant to have the protein that beta carotene is changed into the enzymatic activity of canthaxanthin.

Therefore, think that the ketolase activity is meant the amount of the canthaxanthin of the amount of the beta carotene that transforms by the protein ketolase or formation in special time.

According to it is considered herein that term " wild type " is meant corresponding non-genetic modification Tagetes source plant.

Based on context, term " plant " can be considered to be meant the plants of tagetes species of Tagetes source plant (wild type) or genetic modification of the present invention, or both.

Preferably, think that " wild type " is meant for producing the ketolase activity, increasing, increase and reduce and content astaxanthin increases with reference to plant for hereinafter described ε-cyclase activity for beta cyclase hereinafter described is active for hereinafter described hydroxylase activity under each situation.

Tagetes is marigold, maidenhair, spiceleaf marigold, general woods marigold, Tagetes palmeri, little marigold or bell marigold with reference to plant, it particularly preferably is marigold, very particularly preferably be marigold L., accession number: TAG 72, strain Orangenprinz, can be from IPKGenebank, Corrensstr.3, D-06466 Gatersleben obtains.

The ketolase activity is preferably measured under the following conditions in the plants of tagetes species of genetic modification of the present invention and wild type or in reference to plant:

Measure ketolase activity in the vegetable material based on the method (J.Biol.Chem.272 (10): 6128-6135,1997) of Frazer etc.When having lipid (soybean lecithin) and detergent (sodium taurocholate), utilize the substrate beta carotene and utilize canthaxanthin to measure ketolase activity in the plant extracts.Can measure by HPLC from substrate/product ratio that ketolase is measured.

In this embodiment preferred, to compare with non-genetic modification wild type, the plants of tagetes species of genetic modification of the present invention has the ketolase activity, preferably has the ketolase activity in floral leaf, and therefore preferably can the express transgenic ketolase.

In another preferred embodiment, make in plants of tagetes species, to have the ketolase activity by the gene expression of nucleic acid that causes the coding ketolase.

In this embodiment preferred, preferably import the gene expression of the nucleic acid that causes the coding ketolase in the initial plant of Tagetes by the nucleic acid of the ketolase of will encoding.

By and large, any ketolase gene that is to say any nucleic acid of coding ketolase to can be used in this aspect.

All nucleic acid of mentioning in specification can be, for example, and RNA, DNA or cDNA sequence.

The genome ketolase sequence that contains introne for the eucaryon source, can not express under the situation that corresponding ketolase maybe can not be endowed the ability of expressing corresponding ketolase at the Tagetes host plant, preferably use the nucleotide sequence after having processed, for example corresponding cDNA.

The nucleic acid of coding ketolase is to be selected from following sequence with the example that can be used in the corresponding ketolase of the inventive method:

Haematococcus pluvialis is especially from haematococcus pluvialis Flotow em.Wille (accession number: X86782; Nucleic acid: SEQ ID NO:1, protein s EQ ID NO:2),

Haematococcus pluvialis, NIES-144 (accession number: D45881; Nucleic acid: SEQ ID NO:3, protein s EQ ID NO:4),

Orange Agrobacterium (Agrobacterium aurantiacum) (accession number: D58420; Nucleic acid: SEQ IDNO:5, protein s EQ ID NO:6),

Alcaligenes (Alicaligenes spec.) (accession number: D58422; Nucleic acid: SEQ ID NO:7, protein s EQ ID NO:8),

Paracoccus marcusii (accession number: Y15112; Nucleic acid: SEQ ID NO:9, protein s EQID NO:10),

Synechococcus belongs to (Synechocystis sp.) PC6803 strain (accession number: NP442491; Nucleic acid: SEQ IDNO:11, protein s EQ ID NO:12),

Slowly the knurl of taking root Pseudomonas (Bradyrhizobium sp.) (accession number: AF218415; Nucleic acid: SEQ ID NO:13, protein s EQ ID NO:14),

Nostoc (Nostoc sp.) PCC7120 strain (accession number: AP003592, BAB74888; Nucleic acid: SEQ ID NO:15, protein s EQ ID NO:16),

Point shape beads algae (Nostoc punctiforme) ATTC 29133, nucleic acid: Acc.No.NZ_AABC01000195, base-pair 55,604-55,392 (SEQ ID NO:81); Protein: Acc.No.ZP_00111258 (SEQ ID NO:82) (note) to inferring protein,

Point shape beads algae ATTC 29133, nucleic acid: Acc.No.NZ_AABC01000196, base-pair 140,571-139,810 (SEQ ID NO:83), protein: (SEQ ID NO:84) (do not have and explain), Synechococcus belongs to 8102 kinds of (Synechococcus) WH, nucleic acid: Acc.No.

NZ_AABD01000001, base-pair 1,354,725-1,355,528 (SEQ ID NO:85), protein: Acc.No.ZP 00115639 (SEQ ID NO:86) (note) to inferring protein,

Haematococcus pluvialis (accession number: AF534876, AAN03484; Nucleic acid: SEQ ID NO:97, protein s EQ ID NO:98),

Secondary coccus (Paracoccus) MBIC1143 kind, (accession number: D58420, P54972; Nucleic acid: SEQ ID NO:99, protein s EQ ID NO:100),

Orange shortwave monad (Brevundimonas aurantiaca) (accession number: AY166610, AAN86030; Nucleic acid: SEQ ID NO:101, protein s EQ ID NO:102),

Foam joint ball algae (Nodularia spumigena) NSOR10 (accession number: AY210783, AAO64399; Nucleic acid: SEQ ID NO:103, protein EQ ID NO:104) and

The unusual coccus of radiation hardness (Deinococcus radiodurans) R1 (accession number: E75561, AE001872; Nucleic acid: SEQ ID NO:105, protein: SEQ ID NO:106).

By will from the nucleotide sequence of the amino acid sequence of database or respective opposed translation and above-mentioned sequence and especially with sequence SEQ ID NO:2 and/or 16 comparison homogeneity, can easily find, for example find can be used in the ketolase of the inventive method and other natural example of ketolase gene from the known organism of multiple its genome sequence.

By hybridization technique in a manner known way, can also be easily from above-mentioned nucleotide sequence especially SEQ ID NO:2 and/or 16 from find other natural example of ketolase and ketolase gene from the known organic sequence of multiple its genome sequence.

Hybridization can be at gentle (low strict), or preferably carry out under strict (highly strict) condition.

This type of hybridization conditions is described in, Sambrook for example, J., Fritsch, E.F., Maniatis, T., Molecular Cloning (A Laboratory Manual), second edition, Cold Spring HarborLaboratory Press, 1989,9.31-9.57 page or leaf or Current Protocols in MolecularBiology, John Wiley ﹠amp; Sons, N.Y. (1989), 6.3.1-6.3.6.

For example, that the condition in the washing step can be selected from is low strict (50 ℃ with 2 * SSC) and highly strict (50 ℃, preferably at 65 ℃, with the condition and range of 0.2 * SSC) (pH 7.0 for 20 * SSC:0.3M natrium citricum, 3M sodium chloride) condition restriction.

In addition, the temperature in the washing step can rise to 65 ℃ stringent condition from the temperate condition of 22 ℃ of room temperatures.

Two parameter salinity and temperature can change simultaneously, and also one of these two parameters can be remained unchanged and only another change.In the crossover process, also can use denaturant, for example formamide or SDS.When having 50% formamide, hybridization is preferably carried out at 42 ℃.

Some typical hybridization and the washing step condition is following provides:

(1) hybridization conditions has, for example

(i) 65 ℃, 4 * SSC, or

(ii) 45 ℃, 6 * SSC, or

(iii) 68 ℃, 6 * SSC, 100mg/ml sex change fish sperm DNA, or

(iv) 68 ℃, 6 * SSC, 0.5% SDS, 100mg/ml sex change fragmentation salmon sperm dna, or

(v) 42 ℃, 6 * SSC, 0.5% SDS, 100mg/ml sex change fragmentation salmon sperm dna, 50% formamide, or

(vi) 42 ℃, 50% formamide, 4 * SSC, or

(vii) 42 ℃, 50% (volume/volume) formamide, 0.1% bovine serum albumin(BSA), 0.1% phenanthrene can, 0.1% polyvinylpyrrolidone, 50mM sodium phosphate buffer pH 6.5,750mM NaCl, the 75mM natrium citricum, or

(viii) 50 ℃, 2 * or 4 * SSC (temperate condition), or

(ix) 42 ℃, the 30-40% formamide, 2 * or 4 * SSC (temperate condition).

(2) washing step is 10 minutes under each situation, for example uses

(i) 50 ℃, 0.015M NaCl/0.0015M natrium citricum/0.1% SDS, or

(ii) 65 ℃, 0.1 * SSC, or

(iii) 68 ℃, 0.1 * SSC, 0.5% SDS, or

(iv) 42 ℃, 0.1 * SSC, 0.5% SDS, 50% formamide, or

(v) 42 ℃, 0.2 * SSC, 0.1% SDS, or

(vi) 65 ℃, 2 * SSC (temperate condition).

In the embodiment preferred of the genetically modified plant of Tagetes of the present invention, imported nucleic acid, the protein of these nucleic acid codings contains amino acid sequence SEQ ID NO:2 or by SEQ IDNO:2 is carried out amino acid replacement, insert or disappearance institute calling sequence, homogeneity at these sequences of amino acid levels and sequence SEQ ID NO:2 is at least 20%, be preferably at least 30%, more preferably be at least 40%, more preferably be at least 50%, more preferably be at least 60%, more preferably be at least 70%, more preferably be at least 80%, particularly preferably be at least 90%, and have the enzyme characteristic of ketolase.

Can comprise natural ketolase sequence, as mentioned above, by comparing from other organic sequence, or from sequence SEQ ID NO:2 by the manual change, the natural ketolase sequence that can find of the homogeneity of the synthetic ketolase sequence of having modified by amino acid replacement, insertion or disappearance for example.

In another the inventive method embodiment preferred, imported nucleic acid, the protein of these nucleic acid codings contains amino acid sequence SEQ ID NO:16 or by SEQ ID NO:16 is carried out amino acid replacement, insert or disappearance institute calling sequence, homogeneity at these sequences of amino acid levels and sequence SEQ IDNO:16 is at least 20%, be preferably at least 30%, more preferably be at least 40%, more preferably be at least 50%, more preferably be at least 60%, more preferably be at least 70%, more preferably be at least 80%, particularly preferably be at least 90%, and have the enzyme characteristic of ketolase.

Can comprise natural ketolase sequence, as mentioned above, by comparing from other organic sequence, or from sequence SEQ ID NO:16 by the manual change, the natural ketolase sequence that can find of the homogeneity of the synthetic ketolase sequence of having modified by amino acid replacement, insertion or disappearance for example.

Term " substitutes " and is considered to be meant by one or more amino acid in specification and exchanges one or more amino acid.Preferably, carry out defined conservative exchange, wherein the amino acid that is substituted has similar characteristic to original acid, and for example Glu exchanges to Asp, Gln exchange to Asn, Val exchange to Ile, Leu exchange to Ile, Ser exchange and is Thr.

Disappearance is to replace amino acid by direct key.Preferred disappearance position is the end of polypeptide and the connection between the different proteins domain.

Insertion is that amino acid is imported polypeptide chain, is to replace direct key by one or more amino acid in form.

Homogeneity between two protein is considered to be meant amino acid whose homogeneity in the full length protein under each situation, particularly by using from DNASTAR, inc.Madison, Wisconsin (USA) utilizes the Lasergene software of Clustal method to compare and homogeneity (the Higgins DG that calculates, Sharp PM., on microcomputer, carry out the multisequencing comparison of rapid sensitive, Comput Appl.Biosci.1989 Apr; 5 (2): 151-1), it is as follows that parameter is set:

Multiple ratio is to parameter:

Breach point penalty 10

Notch length point penalty 10

Compare parameter in pairs:

K-tuple (word/word length) 1

Breach point penalty 3

Window 5

Keep diagonal (Diagonals saved) 5

Thought accordingly at the protein that has at least 20% homogeneity with sequence SEQ ID NO:2 or 16 on the amino acid levels and to be meant when with its sequence and sequence SEQ ID NO:2 or 16 relatively the time, especially according to having the program logarithm that above-mentioned parameter is provided with, protein with at least 20% homogeneity.

For example, suitable nucleotide sequence can obtain by the reverse translation of carrying out peptide sequence according to genetic code.

For this respect, preferably use those and the special codon of marigold to select the codon of corresponding to frequent use.Can easily determine the codon selection based on the computer evaluation of other known of plants of tagetes species.

In particularly preferred embodiments, will contain in the nucleic acid importing plants of tagetes species of sequence SEQ ID NO:1.

In another particularly preferred embodiment, the nucleic acid that will contain sequence SEQ ID NO:15 imports in the plants of tagetes species.

In addition, all above-mentioned ketolase genes can produce from the nucleotide structure unit in a manner known way by chemical synthesis, for example pass through the fragment condensation of each overlapping complementary nucleic acid construction unit of double helix.For example, carry out the chemical synthesis of oligonucleotides in known manner by phosphoramidite method (Voet, Voet, second edition, Wiley Press New York, 896-897 page or leaf).The interpolation of synthetic oligonucleotide and utilize the archaeal dna polymerase Klenow fragment to carry out that breach is mended gentle coupled reaction and conventional cloning process is described in (1989) such as Sambrook, Molecular cloning:A laboratory manual, publishing house of cold spring harbor laboratory.

In the inventive method particularly preferred embodiment, use the genetically modified plant of Tagetes, have the highest ketolase expression rate in its petal.

This preferably realizes by expression ketolase gene under the control of flower specific promoter.For example,, with the functional above-mentioned nucleic acid that is connected of flower specific promoter, extensively describe hereinafter in the nucleic acid construct, be imported into plants of tagetes species for this respect.

As initial plant or the preferred especially plants of tagetes species of the genetically modified plant of the present invention, be preferably selected to sow: marigold, maidenhair (they are also referred to as pot marigold), spiceleaf marigold, general woods marigold, Tagetes palmeri, little marigold, lemon marigold, Tagetes signata or bell marigold particularly preferably are marigold or maidenhair.

In preferred embodiments, use to compare and have the hydroxylase activity that improved and/or the genetically modified plant of Tagetes of beta cyclase activity in addition with wild type.

Think that hydroxylase activity is meant the enzymatic activity of hydroxylase.

Think that hydroxylase is meant the protein that has in the enzymatic activity of the optional β that replaces of carotenoid-ionone ring place introducing hydroxyl.

Particularly, think that hydroxylase is meant to have beta carotene is changed into luteole, or canthaxanthin is changed into the protein of the enzymatic activity of astaxanthin.

Therefore, think that hydroxylase activity is meant the beta carotene that transforms by the protein hydroxylase or amount or the luteole of formation or the amount of astaxanthin of canthaxanthin in special time.

Therefore, compare hydroxylase activity with wild type and improve and to be meant and to compare the beta carotene that in special time, transforms with wild type or the amount of canthaxanthin increases, or the amount of luteole that forms or astaxanthin increases.

Preferably, this hydroxylase activity increase is at least 5% of wild type hydroxylase activity, more preferably is at least 20%, more preferably be at least 50%, more preferably be at least 100%, more preferably be at least 300%, even more preferably at least 500%, especially at least 600%.

Think that the beta cyclase activity is meant the enzymatic activity of beta cyclase.

Think that beta cyclase has the protein that the terminal linear residue of lycopene is converted into the enzymatic activity of β-ionone ring.

Particularly, think that beta cyclase is meant to have the protein that gamma carotene is converted into the enzymatic activity of beta carotene.

Therefore, think that the beta cyclase activity is meant in special time the amount of the gamma carotene that is transformed by the protein beta cyclase or the amount of the beta carotene that forms.

Therefore, when comparing with wild type that beta cyclase is active to be improved, the amount of the amount of the gamma carotene that is transformed by the protein beta cyclase or the beta carotene that forms is compared increase with wild type in special time.

Preferably, the increase of this beta cyclase activity is 5% of wild-type beta-cyclase activity at least, more preferably be at least 20%, more preferably be at least 50%, more preferably be at least 100%, more preferably be at least 300%, even more preferably at least 500%, especially at least 600%.

Genetically modified plant of the present invention and wild type or preferably measure under the following conditions with reference to hydroxylase activity in the plant:

(Biochim.Biophys.Acta 1391 (1998), 320-328) activity of method external test hydroxylase according to Bouvier etc.With ferredoxin, ferredoxin-NADP oxidoreducing enzyme, catalase, NADPH and beta carotene,, join in a certain amount of plant extracts with single-or two-galactosylglyceride.

Particularly preferably, according to Bouvier, Keller, d ' Harlingue and Camara (biosynthesis of lutein: from the molecule of the carotenoid hydroxylase enzymes of capsicum (Capsicum annuum L.) and functional characteristic (Biochim.Biophys.Acta 1391 (1998), and 320-328) method is measured hydroxylase activity under the following conditions:

External test carries out in the 0.250ml volume.Measure mixture and contain 50mM potassium phosphate (pH 7.6), 0.025mg spinach ferredoxin, the 0.5 ferredoxin-NADP of unit ⁺Spinach oxidoreducing enzyme, 0.25mM NADPH, 0.010mg beta carotene (being emulsifiable in the 0.1mg Tween 80), 0.05mM be single-and two-galactosylglyceride mixture (1: 1), 1 unit catalase, 200 single-and the plant extracts of two-galactosylglyceride (1: 1), 0.2mg bovine serum albumin(BSA) and different volumes.Reactant mixture was hatched 2 hours at 30 ℃.With organic solvent such as acetone or chloroform/methanol (2: 1), extractive reaction product and measure with HPLC.

Genetically modified plant of the present invention and wild type or preferably measure under the following conditions with reference to beta cyclase activity in the plant:

Activity according to Fraser and Sandmann (Biochem.Biophys.Res.Comm.185 (1) (1992) 9-15) method external test beta cyclase.With kaliumphosphate buffer (pH 7.6), lycopene, pimiento stroma protein, NADP as substrate ⁺, NADPH and ATP join in a certain amount of plant extracts.

Particularly preferably, according to Bouvier, d ' Harlingue and the Camara (analysis of molecules that the carotenoid cyclase suppresses; Arch.Biochem.Biophys.346 (1) (1997) 53-64) method is measured hydroxylase activity under the following conditions:

External test carries out in 250 μ l volumes.Measure plant extracts, 20nM lycopene, 250 μ g pimiento chromoplast stroma proteins, 0.2mM NADP that mixture contains 50mM potassium phosphate (pH 7.6), different amounts ⁺, 0.2mM NADPH and 1mM ATP.Before medium is hatched in adding, immediately NADP/NADPH and ATP are dissolved in the 10ml ethanol with the 1mg Tween 80.30 ℃ of reactions added chloroform/methanol (2: 1) cessation reaction after 60 minutes.The product of being extracted with the HPLC methods analyst.

Another kind utilizes the determination method of radioactive substrates to be described in Fraser and Sandmann (Biochem.Biophys.Res.Comm.185 (1) (1992) 9-15).

Can increase hydroxylase activity and/or beta cyclase activity in several ways, for example express and protein level by closing the inhibition regulatory mechanism, perhaps, increase the gene expression of the nucleic acid of the nucleic acid of coding hydroxylase or the beta cyclase of encoding by comparing with wild type.

Compare with wild type equally, can increase the gene expression of nucleic acid of the nucleic acid or the coding beta cyclase of coding hydroxylase in several ways, for example induce '-hydroxylase gene and/or beta cyclase gene or the nucleic acid of at least one coding hydroxylase or the nucleic acid of at least one coding beta cyclase are imported in the plants of tagetes species by one or more '-hydroxylase genes copies and/or beta cyclase gene copy be that is to say by activator.

The gene expression increase of the nucleic acid of coding hydroxylase and/or beta cyclase also is considered to be meant according to the present invention the manipulation to the expression of plants of tagetes species endogenous hydroxylase and/or beta cyclase.

For example, this can realize by the promoter DNA sequence of modifying coding hydroxylase and/or beta cyclase gene.For example, can realize that by dna sequence dna disappearance or insertion this kind causes the modification that the gene expression rate increases.

As mentioned above, may change the expression of endogenous hydroxylase and/or beta cyclase by the application exogenous irritant.This can that is to say by special physiological conditions by using foreign substance and be undertaken.

And, access endogenous hydroxylase and/or beta cyclase expression of gene altered or that increased by using in non-conversion plant non-existent adjusting albumen and this gene promoter interaction energy.

The chimeric protein that this kind conditioning agent can be made up of DNA-binding structural domain and transcription activator domain for example is described in the conditioning agent among the WO 96/06166.

In preferred embodiments, by with the nucleic acid of at least one coding hydroxylase or import the gene expression of the nucleic acid of the gene expression of the nucleic acid that increases the coding hydroxylase in the plants of tagetes species and/or coding beta cyclase by nucleic acid with at least one coding beta cyclase.

For this respect, can use any '-hydroxylase gene or any beta cyclase gene in principle, that is to say the nucleic acid of any coding hydroxylase and the nucleic acid of the beta cyclase of encoding arbitrarily.

Genome hydroxylase that contains introne or beta cyclase nucleotide sequence for the eucaryon source, do not have maybe can not be endowed under the situation of the ability of expressing corresponding hydroxylase or beta cyclase at host plant, preferably use the nucleotide sequence of processing through in advance, such as corresponding cDNA.

An example of '-hydroxylase gene is from haematococcus pluvialis (accession number AX038729, WO 0061764); (nucleic acid: SEQ ID NO:17, protein: the nucleic acid of coding hydroxylase SEQ ID NO:18).

Also can use hydroxylase with following accession number:

|emb|CAB55626.1、CAA70427.1、CAA70888.1、CAB55625.1、AF499108_1、AF315289_1、AF296158_1、AAC49443.1、NP_194300.1、NP_200070.1、AAG10430.1、CAC06712.1、AAM88619.1、CAC95130.1、AAL80006.1、AF162276_1、AAO53295.1、AAN85601.1、CRTZ_ERWHE、CRTZ_PANAN、BAB79605.1、CRTZ_ALCSP、CRTZ_AGRAU、CAB56060.1、ZP_00094836.1、AAC44852.1、BAC77670.1、NP_745389.1、NP_344225.1、NP_849490.1、ZP_00087019.1、NP_503072.1、NP_852012.1、NP_15929.1、ZP_00013255.1

Particularly preferred in addition hydroxylase is hydroxylase (accession number the Y14809) (nucleic acid: SEQ ID NO:107 from tomato; Protein: SEQ ID NO.108).

The example of beta cyclase gene is:

Coding is from nucleic acid (accession number the X86452) (nucleic acid: SEQ ID NO:19, protein: SEQ ID NO:20) of the beta cyclase of tomato.

Also can use beta cyclase with following accession number:

S66350 lycopene beta cyclase (EC 5.5.1.-)-tomato

CAA60119 lycopene synthase [capsicum (Capsicum annuum)]

S66349 lycopene beta cyclase (EC 5.5.1.-)-common tobacco

CAA57386 lycopene cyclase [tobacco (Nicotiana tabacum)]

AAM21152 lycopene beta cyclase [sweet orange (Citrus sinensis)]

AAD38049 lycopene cyclase [grape fruit (Citrus x paradisi)]

AAN86060 lycopene cyclase [satsuma orange (Citrus unshiu)]

AAF44700 lycopene beta cyclase [sweet orange]

AAK07430 lycopene beta cyclase [John Burroughs carry receive adonis amurensis (Adonispalaestina)]

AAG10429 beta cyclase [marigold]

The AAA81880 lycopene cyclase

The AAB53337 lycopene beta cyclase

AAL92175 β-lycopene cyclase [palace lantern lily (Sandersonia aurantiaca)]

CAA67331 lycopene cyclase [daffodil (Narcissus pseudonarcissus)]

AAM45381 beta cyclase [marigold]

AAO18661 lycopene beta cyclase [corn (Zea mays)]

The lycopene beta cyclase [edible tomato (Lycopersiconesculentum)] that the AAG21133 chromoplast is special

AAF18989 lycopene beta cyclase [carrot (Daucus carota)]

ZP_001140 infers protein [the former green coccus in ocean (Prochlorococcus marinus) MIT9313 strain]

ZP_001050 infers protein [ocean former green coccus Maas moral subspecies CCMP1378 strain]

ZP_001046 infers protein [ocean former green coccus Maas moral subspecies CCMP1378 strain]

ZP_001134 infers protein [ocean former green coccus MIT9313 strain]

ZP_001150 infers protein [Synechococcus belongs to 8102 kinds of WH]

AAF10377 lycopene cyclase [the unusual coccus of radiation hardness (Deinococcusradiodurans)]

BAA29250 393aa length is inferred protein [Huo Shi fireball bacterium (Pyrococcus horikoshii)]

BAC77673 lycopene beta-single cyclase [marine products bacterium P99-3]

AAL01999 lycopene cyclase [Flavobacterium (Xanthobacter) Py2 kind]

ZP_000190 infers protein [the orange green der Pilz (Chloroflexus aurantiacus) that deflects]

ZP_000941 infers protein [Novosphingobium aromaticivorans]

AAF78200 lycopene cyclase [knurl of taking root slowly Pseudomonas ORS278]

BAB79602 crtY[reticulate millettia pantoea agglomerans (Pantoea agglomerans pv.Milletiae)]

CAA64855 lycopene cyclase [streptomyces griseus (Streptomyces griseus)]

AAA21262 lycopene cyclase [pantoea agglomerans (Pantoea agglomerans)]

C37802 crtY protein-Erwinia uredovora (Erwinia uredovora)

BAB79602 crtY [reticulate millettia pantoea agglomerans]

AAA64980 lycopene cyclase [pantoea agglomerans]

The AAC44851 lycopene cyclase

BAA09593 lycopene cyclase [secondary coccus MBIC1143 kind]

ZP_000941 infers protein [Novosphingobium aromaticivorans]

CAB56061 lycopene beta cyclase [Paracoccus marcusii]

BAA20275 lycopene cyclase [long bacillus rubidus (Erythrobacter longus)]

ZP_000570 infers protein [having a liking for thermophilic actinomycete (Thermobifida fusca)]

ZP_000190 infers protein [the orange green der Pilz that deflects]

AAK07430 lycopene beta cyclase [John Burroughs carry receive adonis amurensis]

CAA67331 lycopene cyclase [daffodil]

The AAB53337 lycopene beta cyclase

BAC77673 lycopene beta-single cyclase [marine products bacterium P99-3]

In addition, particularly preferred beta cyclase is from the special beta cyclase of tomato plastid (AAG21133) (nucleic acid: SEQ ID No.109; Protein: SEQ ID No.110).

Therefore, in this embodiment preferred, compare, in the preferred Tagetes genetically modified plants of the present invention, a '-hydroxylase gene and/or beta cyclase gene are arranged at least additionally with wild-type plant.

For example, genetically modified in preferred embodiments plant has the exogenous nucleic acid of at least a coding hydroxylase, or the endogenous nucleic acid of at least two kinds of coding hydroxylases, and/or the exogenous nucleic acid of at least a coding beta cyclase, or the endogenous nucleic acid of at least two kinds of coding beta cyclases.

Preferably, in above-mentioned embodiment preferred, as '-hydroxylase gene, the nucleic acid coding that uses contains amino acid sequence SEQ ID NO:18 or by this sequence being carried out amino acid replacement, insertion or lacking the protein of resulting sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ IDNO:18 is at least 30%, preferably be at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and this protein has the enzyme characteristic of hydroxylase.

For example, as mentioned above, by carrying out the homology comparison with SEQ ID.NO:18, can easily find other hydroxylase and '-hydroxylase gene from the known organism of several genes group sequence from the amino acid sequence of database or corresponding reverse translation nucleotide sequence.

For example, as mentioned above, by hybridization and round pcr in a manner known way, also can easily find other hydroxylase and '-hydroxylase gene from sequence SEQ ID NO:17 is initial from multiple organism from its genome sequence the unknown.

In another particularly preferred embodiment, nucleic acid is imported organism to increase hydroxylase activity, wherein the protein of nucleic acid coding contains the amino acid sequence of the hydroxylase of sequence SEQ ID NO:18.

For example, can obtain suitable nucleotide sequence according to the reverse translation of genetic code by peptide sequence.

For this respect, preferably use those to select the frequent codon that uses according to the special codon of plant.Select based on can determine codon fast to the computer evaluation of relevant other known of organism.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:17.

In above-mentioned embodiment preferred, as the beta cyclase gene, the preferred nucleic acid coding that uses contains amino acid sequence SEQ ID NO:20 or by this sequence being carried out amino acid replacement, insertion or lacking the protein of resulting sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ IDNO:20 is at least 30%, preferably be at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and this protein has the enzyme characteristic of beta cyclase.

For example, as mentioned above, by carrying out the homology comparison with SEQ ID.NO:20, can easily find other beta cyclase and beta cyclase gene from the known organism of several genes group sequence from the amino acid sequence of database or corresponding reverse translation nucleotide sequence.

For example, by hybridization and round pcr in a manner known way, also can be easily initial from finding other beta cyclase and beta cyclase gene from the multiple organism of its genome sequence the unknown from sequence SEQ ID NO:19.

In another particularly preferred embodiment, nucleic acid is imported organism to increase the beta cyclase activity, wherein the protein of nucleic acid coding contains the amino acid sequence of the beta cyclase of sequence SEQ ID NO:20.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:19.

In addition, all above-mentioned '-hydroxylase genes or beta cyclase gene can produce from the nucleotide structure unit in a manner known way by chemical synthesis, for example pass through the fragment condensation of each overlapping complementary nucleic acid construction unit of double helix.For example, carry out the chemical synthesis of oligonucleotides in known manner by phosphoramidite method (Voet, Voet, second edition, WileyPress New York, 896-897 page or leaf).The interpolation of synthetic oligonucleotide and utilize the archaeal dna polymerase Klenow fragment to carry out that breach is mended gentle coupled reaction and conventional cloning process is described in (1989) such as Sambrook, Molecular cloning:Alaboratory manual, publishing house of cold spring harbor laboratory.

In addition, in another embodiment preferred of this method, compare with wild type, plants of tagetes species has the ε-cyclase activity of reduction.

Think that ε-cyclase activity is meant the enzymatic activity of ε-cyclase.

Think that ε-cyclase is meant to have the protein that the terminal linear residue of lycopene is converted into the enzymatic activity of ε-ionone ring.

Therefore think that ε-cyclase particularly is meant to have the protein that lycopene is converted into the enzymatic activity of δ-carrotene.

Therefore, think that ε-cyclase activity is meant in special time the amount of the lycopene that is transformed by protein ε-cyclase or the amount of δ-carrotene of forming.

Therefore, when ε-cyclase activity is compared with wild type when reducing, in special time, compare the amount of the lycopene that transforms by protein ε-cyclase or the amount of δ-carrotene of forming reduces with wild type.

Based on different cell biology mechanism, think that preferably ε-the cyclase activity that reduces is meant part or whole basically function that suppresses or block ε-cyclase in plant cell, plant or its part, tissue, organ, cell or the seed.

For example, compare, can reduce ε-cyclase activity in the plant by the amount of ε-cyclase protein matter or the amount of ε-cyclase mRNA in the reduction plant with wild type.Therefore, compare with wild type, can be directly or the quantitative determination of the amount by measuring ε-cyclase protein matter in the plant of the present invention or ε-cyclase mRNA compare the ε-cyclase activity that has reduced with wild type.

The reduction of ε-cyclase activity comprises that ε-cyclase is reduced to and lacks ε-cyclase (that is to say and lack detectable ε-cyclase activity or lack the detectable ε-cyclase of immunology) basically fully on amount.Preferably, particularly preferably be to spend middle ε-cyclase activity (or amount of the amount of ε-cyclase protein matter or ε-cyclase mRNA) to compare reduction at least 5% in the plant with wild type, more preferably reduce at least 20%, more preferably reduce at least 50%, more preferably reduce by 100%.Particularly, " reduction " also be meant and lack ε-cyclase activity (or ε-cyclase protein matter or ε-cyclase mRNA) fully.

In the genetically modified plant of the present invention and wild type or preferably measure under the following conditions with reference to ε-cyclase activity in the plant:

According to Fraser and Sandmann (Biochem.Biophys.Res.Comm.185 (1) (1992) 9-15) method can external test ε-cyclase activity, during mensuration, will be as potassium phosphate (pH 7.6), lycopene, pimiento stroma protein, the NADP of buffer solution as substrate ⁺, NADPH and ATP join in a certain amount of plant extracts.

Particularly preferably, according to Bouvier, d ' Harlingue and the Camara (analysis of molecules that the carotenoid cyclase suppresses; Arch.Biochem.Biophys.346 (1) (1997) 53-64) method is measured the genetically modified plant of the present invention and wild type or with reference to the activity of ε-cyclase in the plant:

External test carries out in the 0.25ml volume.Measure plant extracts, 20nM lycopene, 0.25mg pimiento stroma protein, 0.2mM NADP that mixture contains 50mM potassium phosphate (pH 7.6), different amounts ⁺, 0.2mM NADPH and 1mM ATP.Before hatching medium, adding immediately NADP/NADPH and ATP are dissolved in 0.01ml ethanol with the 1mg Tween 80.After 60 minutes, add chloroform/methanol (2: 1) cessation reaction 30 ℃ of reactions.Analyze the product of being extracted in the chloroform with HPLC.

Utilize the another kind of determination method of radioactive substrates to be described in Fraser and Sandmann (Biochem.Biophys.Res.Comm.185 (1) (1992) 9-15).Another kind of analytic approach is described in Beyer, Kr_ncke and the Nievelstein (mechanism of lycopene isomerase in the daffodil chromoplast/cyclase reaction; J.Biol.Chem.266 (26) (1991) 17072-17078).

Preferably reduce the activity of ε-cyclase in the plant by at least a following method:

A) import at least a double-stranded ε-cyclase RNA sequence, be also referred to as ε-cyclase-dsRNA hereinafter, or guarantee an expression cassette or a plurality of expression cassette of its expression.Comprise those wherein ε-cyclase-dsRNA at the method for ε-cyclase gene (that is to say genomic dna sequence) or ε-cyclase transcript (that is to say the mRNA sequence) such as promoter sequence

B) import at least a ε-cyclase antisense RNA sequence, be also referred to as ε-cyclase-antisense RNA hereinafter, or guarantee the expression cassette of its expression.Comprise those wherein ε-cyclase-antisense RNA at the method for ε-cyclase gene (that is to say genomic dna sequence) or ε-cyclase transcript (that is to say the RNA sequence).Also comprise α-different head (anomeric) nucleotide sequence

C) import the ε-cyclase antisense RNA of at least a and ribozyme combination or guarantee the expression cassette of its expression

D) import at least a ε-cyclase and have a mind to RNA sequence, be also referred to as ε-cyclase-have a mind to RNA hereinafter, inducing common inhibition, or import the expression cassette of guaranteeing its expression

E) import at least aly, perhaps import the expression cassette of guaranteeing its expression at ε-cyclase gene, RNA or protein DNA-or protein-binding factor

F) import at least a nucleic acid sequence that causes ε-cyclisation ribozyme degraded, perhaps import the expression cassette of guaranteeing its expression

G) import at least a construct to produce ε-cyclase gene afunction, such as producing the displacement of terminator codon or reading frame, for example by in ε-cyclase gene, producing insertion, disappearance, inversion or sudden change.Preferably, inserting described ε-cyclase gene or implanting needle by homologous recombination with target produces the mode of the sequence-specific nuclease of ε-cyclase gene sequence and knocks out sudden change.

Other known for those skilled in the art method also can be used in the content of the present invention to reduce ε-cyclase or its activity or function.For example, import ε-cyclase dominant (dominantnegative) variant or guarantee that the expression cassette of its expression may also be favourable.Each method can both cause the ε-amount of cyclase protein matter, the amount of mRNA and/or active the reduction in these methods.Also can these methods of applied in any combination.Additive method also is known for those skilled in the art and can comprises the transhipment that stops or suppress processing, ε-cyclase or its mRNA of ε-cyclase, suppressing ribosomes adheres to, suppress the RNA montage, induce ε-cyclase RNA-digestive enzyme and/or suppress the translation extension or the termination translation.

Various preferable methods can be described in by exemplary embodiment hereinafter:

A) import double-stranded ε-cyclase RNA sequence (ε-cyclase-dsRNA) with double-stranded RNA mode (" double-stranded RNA interference "; DsRNAi) method of carrying out gene regulation is known and is described in such as (2000) Plant Mol Biol43:401-415 such as Matzke MA; (1998) Nature 391:806-811 such as Fire A.; WO 99/32619; WO 99/53050; WO 00/68374; WO 00/44914; WO 00/44895; WO 00/49035 or WO 00/63364.Therefore be described in the step in the quoted passage reported and method specific reference in the application as a reference.

According to the present invention, think that " double stranded RNA sequence " is meant because complementary series, in theory as according to the base pairing rules of Watson and Crick and/or in fact as based on hybrid experiment, can be external and/or body in form one or more RNA sequence of double-stranded RNA structure.

The formation of the known double-stranded RNA structure of those skilled in the art is a kind of equilibrium state.Preferably, the ratio of duplex molecule and the corresponding form of unwinding was at least 1: 10, was preferably 1: 1, was 5: 1 particularly preferably, was most preferably 10: 1.

Think that double-stranded ε-cyclase RNA sequence or ε-cyclase-dsRNA preferably are meant to have the RNA molecule that contains the duplex structure zone and contain following nucleotide sequence in this zone:

A) with the intrinsic ε-cyclase of plant to the small part transcript identical and/or

B) identical with the intrinsic ε-cyclase of plant to the small part promoter sequence.

In the methods of the invention, in order to reduce ε-cyclase activity, preferably such RNA is imported plant, described RNA has the zone that contains duplex structure and contain following nucleotide sequence in this zone

Think that term " ε-cyclase transcript " is meant the part of transcribing of ε-cyclase gene, wherein except the sequence of coding ε-cyclase, also contain such as non-coding sequence, as UTRs.

Think the RNA of " with the intrinsic ε-cyclase of plant identical " to the small part promoter sequence preferably be meant with ε-cyclase promoter sequence that is to say the RNA sequence that corresponding RNA sequence is identical to the theoretical transcript of small part.

Think that intrinsic ε-cyclase " part " promoter sequence of the intrinsic ε-cyclase of plant " part " transcript or plant is meant the partial sequence from several base-pairs to the as many as full sequence in transcript or the promoter sequence.By those skilled in the art by the normal experiment optimal length of determining section sequence easily.

Usually, the length of partial sequence is at least 10 bases and maximum 2kb, be preferably at least 25 bases and maximum 1.5kb, particularly preferably be at least 50 bases and maximum 600 bases, very particularly preferably be at least 100 bases and maximum 500 bases, be most preferably at least 200 bases or at least 300 bases and maximum 400 bases.

Preferably, the mode that does not reduce with the activity that obtains high as far as possible specificity and other enzyme (not expecting its active reduction) is found out partial sequence.Therefore the partial sequence that is chosen in non-existent ε-cyclase part transcript in other activity and/or the ε-cyclase promoter sequence for ε-cyclase-dsRNA partial sequence is favourable.

Therefore, in particularly preferred embodiments, ε-cyclase-dsRNA contains the identical sequence of the part ε intrinsic with plant-cyclase transcript and contains 5 ' terminal or 3 ' end of the intrinsic coding ε-cyclase nucleic acid of plant.Especially, transcript 5 ' or 3 ' non-translational region are suitable for producing the selectivity duplex structure.

The invention further relates to double stranded rna molecule (dsRNA molecule), when it is imported plant (or its cell, tissue, organ or propagating materials), cause that ε-cyclase reduces.

(ε-cyclase-dsRNA) express, double stranded rna molecule preferably comprises for reducing ε-cyclase

A) contain at least a identical with at least a portion of " having a mind to " RNA-ε-cyclase transcript basically ribonucleotide acid sequence " having a mind to " RNA chain and

B) basically preferably fully and " antisense " RNA chain of " having a mind to " the chain complementation of the RNA a).

For transforming plant, preferably use the nucleic acid construct that is imported in the plant and in plant, is transcribed into ε-cyclase-dsRNA with ε-cyclase-dsRNA.

Therefore, the invention still further relates to and to be transcribed into following nucleic acid construct

B) basically preferably fully and " antisense " RNA chain of " having a mind to " the chain complementation of the RNA a).These nucleic acid constructs are also referred to as expression cassette or expression vector hereinafter.

About the dsRNA molecule, think that ε-cyclase nucleotide sequence or corresponding transcript preferably are meant the sequence according to SEQ ID NO:38 or its part.

" substantially the same " meaning is to compare with ε-cyclase target sequence, and the dsRNA sequence also can have insertion, disappearance and single point mutation, yet causes that still expression effectively reduces.Preferably, " having a mind to " chain of inhibition dsRNA and ε-cyclase gene to small part " is had a mind to " between the rna transcription basis or the homology between " antisense " chain and the ε-cyclase gene complementary strand is at least 75%, be preferably at least 80%, very particularly preferably be at least 90%, be most preferably 100%.

Reducing for causing that ε-cyclase is expressed effectively, is not to have 100% homogeneity between definitely necessary dsRNA and the ε-cyclase gene transcript.So owing to may have genetic mutation, polymorphism or evolutionary divergence, the method that the tolerance sequence departs from is favourable.Therefore, for example, it is possible utilizing dsRNA by a kind of organism ε-initial generation of cyclase sequence to suppress that ε in the another kind of organism-cyclase expresses.For this purpose, dsRNA preferably contains the sequence area of ε-cyclase gene transcript corresponding to conserved region.Can easily obtain described conserved region by the sequence comparison.

In addition, the dsRNA of " substantially the same " also can be defined as the nucleotide sequence that can hybridize (hybridizing 12-16 hour as 50 ℃ or 70 ℃) with a part of ε-cyclase gene transcript in 400mM NaCl, 40mM PIPES pH 6.4,1mM EDTA.

" complementary basically " meaning is meant with the complementary strand of " having a mind to " RNA chain and compares that " antisense " RNA chain also can have insertion, disappearance and single point mutation.Preferably, the homology between the complementary strand of " antisense " RNA chain and " having a mind to " RNA chain is at least 80%, is preferably at least 90%, very particularly preferably is at least 95%, is most preferably 100%.

In another embodiment, ε-cyclase-dsRNA comprises

A) contain at least a basically with ε-cyclase gene promoter region " have a mind to " the identical ribonucleotide acid sequence of this at least a portion of rna transcription " having a mind to " RNA chain and

B) basically preferably fully and " antisense " RNA chain of " having a mind to " the chain complementation of the RNA a).The corresponding nucleic construct that is preferably used for transforming plant comprises

A) identical with at least a portion of ε-cyclase gene promoter region basically " having a mind to " DNA chain and

B) basically preferably fully and " antisense " DNA chain of " having a mind to " the chain complementation of the DNA a).Preferably, think that ε-cyclase promoter region is meant according to sequence SEQ ID NO:47 or its partial sequence.

Particularly,, particularly preferably use following partial sequence in order to produce the ε-cyclase-dsRNA sequence that reduces ε-cyclase activity for marigold:

SEQ ID NO:40: the fragment intentionally of ε-cyclase 5 '-stub area

SEQ ID NO:41: the antisense fragment of ε-cyclase 5 '-stub area

SEQ ID NO:42: the fragment intentionally of ε-cyclase 3 '-stub area

SEQ ID NO:43: the antisense fragment of ε-cyclase 3 '-stub area

The fragment intentionally of SEQ ID NO:47: ε-cyclase promoter

The antisense fragment of SEQ ID NO:48: ε-cyclase promoter

DsRNA can be made up of one or more polybribonucleotide chain.Obviously, in order to realize identical purpose, also can be with in multiple different the dsRNA molecule transfered cell or organism, wherein each dsRNA molecule all contains a kind of ribonucleotide acid sequence part defined above.

Can be by the independent RNA start of chain of two complementations or preferably form double-stranded dsRNA structure by the RNA start of chain of self complementation.In this case, " have a mind to " RNA chain and " antisense " RNA chain preferably with the form of oppositely " repetition " covalent bond each other.

For example, described in WO 99/53050, dsRNA also can include hairpin structure, by with catenation sequence (" joint "; Introne for example) connects " having a mind to " and " antisense " chain.The dsRNA structure of self complementation is preferred, and this is because they only need the complementary RNA chain of molar ratios such as expressing a RNA sequence and always contain.Preferably, catenation sequence is that introne is (for example from the introne of potato ST-LS1 gene; (1990) Mol Gen Genet220 (2) such as Vancanneyt GF: 245-250).

The nucleotide sequence of coding dsRNA can contain other element, for example transcription stop signals or polyadenylation signal.

Yet, if dsRNA at ε-cyclase promoter sequence, does not preferably contain transcription stop signals or polyadenylation signal.This can be in nucleus retention dsRNA and stop dsRNA in whole strain plant, distribute (" diffusion ").

If desire imports a cell or plant together with two chains of dsRNA, can for example implement in the following manner:

A) with the carrier transformant or the plant that contain two kinds of expression cassettes,

B) use two kinds of carrier cotransformation cells or plant, a kind of carrier contains the expression cassette with " having a mind to " chain, and another kind of carrier contains the expression cassette with " antisense " chain.

C) the different plant lines hybridization of two strains, a strain contains the expression cassette with " having a mind to " chain, and another strain contains the expression cassette with " antisense " chain.

The formation of RNA two strands can be in the extracellular or is initial in cell.

In vivo or externally all can synthesize dsRNA.Therefore, the dna sequence dna of coding dsRNA can be placed at expression cassette under at least one hereditary control element (for example promoter) control.Polyadenylation is optional, and is same, and the element of initial translation neither exist.Preferably, the expression cassette of ε-cyclase dsRNA is present on transformation construct or the conversion carrier.

In particularly preferred embodiments, by under the functional control of flower specific promoter, particularly preferably be under the described promoter of sequence SEQ ID NO:28 or its functional equivalent are partly controlled, from the initial expression of expression construct dsRNA.

Therefore, preferably will the encode expression cassette of ε-cyclase-dsRNA " antisense " and/or " having a mind to " chain or the expression cassette of coding dsRNA self complementary strand is inserted into conversion carrier and uses following method that carrier is imported in the plant cell.For the inventive method, stable to be inserted into genome be favourable.

Can be with dsRNA so that can there be the amount of a copy to import in each cell at least.Higher as required amount (for example each cell at least 5,10,100,500 or 1000 copies) can cause more high efficiency reduction.

B) the antisense RNA sequence (ε-cyclase-antisense RNA) of importing ε-cyclase

Reduce certain method of protein by " antisense " technology and repeatedly be described in plant (Sheehy etc. (1988) Proc Natl Acad Sci USA 85:8805-8809; US 4,801, and 340; (1990) FEBS Lett 268 (2) such as Mol JN: 427-430).The cell mRNA of antisense RNA molecule and the coding ε-cyclase that will be reduced and/or genomic DNA hybridization or combine.This can suppress transcribing and/or translating of ε-cyclase.In a usual manner by the stable two strands of formation or under the situation that is genomic DNA, by hybridizing in the zanjon that antisense nucleic acid molecule is incorporated into genomic DNA double-stranded DNA spiral with special interaction.

According to the basepairing rule of Watson and Crick, utilize the nucleotide sequence of this ε-cyclase of coding, for example, can access ε-cyclase-antisense RNA according to the nucleotide sequence of SEQ ID NO:38.The full length mRNA complementation that ε-cyclase-antisense RNA can be transcribed with ε-cyclase, can be confined to the code area complementation or only by forming with the complementary oligonucleotides of the coding of mRNA or non-coding sequence part.For example, this oligonucleotides can with the regional complementarity that contains ε-cyclase translation initiation part.ε-cyclase-antisense RNA length can be for example 5,10,15,20,25,30,35,40,45 or 50 nucleotides, but also may be longer and contain at least 100,200,500,1000,2000 or 5000 nucleotides.In the context of the inventive method, ε-cyclase-antisense RNA preferably is expressed in the target cell with recombination form.

In particularly preferred embodiments, antisense RNA particularly preferably is under the control of the described promoter of SEQ ID NO:28 or its functional equivalent part by the initial expression of expression construct under the functional control of flower specific promoter.

Described expression cassette can be the part of transformation construct or conversion carrier, perhaps can import by cotransformation.

In the embodiment that is more preferably, can be by complementary with ε-cyclase gene control region (for example ε-cyclase promoter and/or enhancer) and suppress the expression of ε-cyclase with the nucleotide sequence of dna double spiralization triple-helix structure, thus transcribing of ε-cyclase gene reduced.Corresponding method (Helene C (1991) Anticancer Drug Res 6 (6): 569-84 has been described; (1992) Ann NY Acad Sci 660:27-36 such as Helene C; Maher LJ (1992) Bioassays 14 (12): 807-815).

In another embodiment, ε-cyclase-antisense RNA can be a α-different nucleic acid.This type of α-different nucleic acid molecules and complementary RNA form special double-stranded crossbred, and be wherein opposite with the β-nucleic acid of routine, its two chain directions parallel ((1987) Nucleic Acids Res15:6625-6641 such as Gautier C).

C) import the ε-cyclase-antisense RNA that combines with ribozyme

Advantageously, above-mentioned antisense strategy can combine with the ribozyme method.Catalysis RNA molecule or ribozyme can be designed and be applicable to any purpose target RNA and at special site fracture phosphodiester backbone, this can functional inactivation target RNA (Tanner NK (1999) FEMS Microbiol Rev23 (3): 257-275).Ribozyme self is not modified as a result, but other targets RNA molecule that can rupture in a similar manner, this has just given the characteristic of its enzyme.Ribozyme sequence is integrated with " antisense " RNA and given these " antisenses " RNA this kind of enzyme sample RNA cutting characteristic clearly, and therefore increased the efficient of its inactivation target RNA.The generation and the purposes (especially seeing (1988) Nature 334:585-591 such as Haseloff) of corresponding ribozyme " antisense " RNA molecule have been described; Haselhoff and Gerlach (1988) Nature 334:585-591; (1992) EMBO J11 (4): 1525-1530 such as Steinecke P; (1996) Mol Gen Genet.250 (3) such as de Feyter R: 329-338).

In this kind mode, ribozyme (" hammerhead shape " ribozyme for example; Haselhoff and Gerlach (1988) Nature 334:585-591) can be used in catalytic cutting with the ε-cyclase mRNA that is reduced, thereby and stop its translation.The ribozyme technology can improve the efficient of antisense strategy.The method that expression is used for reducing the ribozyme of some protein is described in (EP 0 291 533, EP 0 321 201, EP 0 360 257).Describe ribozyme in the plant cell equally and expressed ((1992) EMBO J 11 (4): 1525-1530 such as Steinecke P; (1996) Mol Gen Genet.250 (3) such as de Feyter R: 329-338).For example, described in " Steinecke P; ribozyme; Methods in Cell Biology 50; editors such as Galbraith, Academic Press, Inc. (1995); 449-460 page or leaf ", by the secondary structure of ribozyme and target RNA being calculated and can being determined suitable target sequence and ribozyme ((1992) Plant Mol Biol.18 (2): 353-361 such as Bayley CC by its interaction; (1994) Mol Gen Genet.242 (6) such as Lloyd AM and Davis RW: 653-657).For example, can make up tetrahymena (Tetrahymena) the L-19 IVS RNA derivative (also see US 4,987,071 and US 5,116,742) in zone with the ε-cyclase mRNA complementation that suppresses with desire.In addition, by also can determine this type of ribozyme (Bartel D and Szostak JW (1993) Science 261:1411-1418) from different ribozyme library screening methods.

D) importing ε-cyclase has a mind to RNA sequence (ε-cyclase-have a mind to RNA) and expresses the common inhibition that ε-cyclase RNA sequence (or its part) can cause corresponding ε-cyclase gene to induce common inhibition to have a mind to direction.Can reduce or close its expression with the expression of RNA intentionally of endogenous ε-cyclase gene homology, describe similar manner (Jorgensen etc. (1996) the Plant Mol Biol 31 (5): 957-973 that is used for the antisense method; Goring etc. (1991) Proc Natl Acad SciUSA 88:1770-1774; Smith etc. (1990) Mol Gen Genet 224:447-481; Napoli etc. (1990) Plant Cell 2:279-289; (1990) Plant Cell 2:291-99 such as Van der Krol).In this article, the construct that is imported can be represented will reduce whole or homeologous gene only.May not need translation.Described this technology has been applied to plant ((1990) Plant Cell 2:279-289 such as Napoli for example, US 5,034, in 323).

Preferably, utilize basically with coding ε-cyclase to the identical sequence of small part nucleotide sequence, for example according to the nucleotide sequence of SEQ ID NO:38, realize suppressing altogether.Preferably, to such an extent as to selection ε-cyclase-translation of ε-cyclase or its part can not take place in RNA intentionally.Therefore, for example, can select 5 '-non-translational region or 3 '-non-translational region, the ATG initiation codon of perhaps also can deleting or suddenly change.

E) implanting needle to ε-cyclase gene, RNA or protein DNA-or protein-binding factor utilize specific DNA-binding factor, for example utilize the zinc finger transcription factor type factor, also may reduce ε-cyclase and express.These factors are incorporated into the endogenous target gene genome sequence and list, and preferably are incorporated into control region, and cause that expression reduces.Correlation technique ((2001) J Biol Chem 276 (31): the 29466-78 such as Dreier B that produces the corresponding factor has been described; (2000) J Mol Biol303 (4): 489-502 such as Dreier B; (2000) Proc Natl Acad Sci USA97 (4): 1495-1500 such as Beerli RR; (2000) J Biol Chem 275 (42): 32617-32627 such as Beerli RR; Segal DJ and Barbas CF 3rd. (2000) Curr Opin Chem Biol4 (1): 34-39; KangJS and Kim JS (2000) J Biol Chem 275 (12): 8742-8748; (1998) Proc Natl Acad Sci USA 95 (25): 14628-14633 such as Beerli RR; (1997) Proc NatlAcad Sci USA 94 (8): 3616-3620 such as Kim JS; Klug A (1999) J Mol Biol 293 (2): 215-218; (1998) Adv Drug Deliv Rev 30 (1-3): 23-31 such as Tsai SY; (2000) Proc Natl Acad Sci USA 97 (8): 3930-3935 such as Mapp AK; (1997) Int JBiochem Cell Biol 29 (12): 1371-1387 such as Sharrocks AD; (2000) J Biol Chem275 (43) such as Zhang L: 33850-33860).

Utilize any purpose fragment of ε-cyclase gene can select these factors.Preferably, this part is in promoter region.Yet, suppressing for gene, this part also can be positioned at coding extron or introne zone.

In addition, the factor of inhibition ε-cyclase self also can transfered cell.These protein-binding factors can be for example fit (aptamer) (Famulok M and Mayer G (1999) Curr TopMicrobiol Immunol 243:123-36) or antibody or antibody fragment or single-chain antibody.Preparation to these factors is described ((1992) Biotechnology (NY) 10 (7): 790-794 such as Owen M; (1997) Curr Opin Biotechnol 8 (4): 411-416 such as Franken E; Whitelam (1996) Trend Plant Sci 1:268-272).

F) import nucleic acid sequence and the expression construct that causes ε-cyclisation ribozyme degraded

By virus expression systems is imported plant, induce special ε-cyclisation ribozyme degraded can realize that also ε-cyclase expression reduces (Amplikon; (1999) Plant J20 (3) such as Angell SM: 357-362).Use the viral vectors nucleotide sequence that has homology with the ε-cyclase transcript that will reduce, these systems are also referred to as " VIGS " (gene silencing of virus induction) import plant.Close then and transcribe, infer that this passes through the plant defense mechanism mediation at virus.Corresponding techniques and method ((2001) Plant J 25 (2): 237-45 such as Ratcliff F has been described; Fagard M and Vaucheret H (2000) Plant Mol Biol 43 (2-3): 285-93; (1998) Proc Natl Acad Sci USA 95 (22): 13079-84 such as Anandalakshmi R; Ruiz MT (1998) Plant Cell10 (6): 937-46).

Preferably, utilize basically with coding ε-cyclase for example realize the minimizing of VIGS-mediation to the identical sequence of small part nucleotide sequence according to the nucleotide sequence of SEQ ID NO:1.

G) import the construct that produces ε-cyclase gene afunction or function reduction

Those skilled in the art know in a large number the method about genome sequence can be modified by specificity.Particularly, these methods for example comprise producing by the target reorganization and knock out sudden change, for example produce terminator codon, reading frame displacement etc. (Hohn B and Puchta H (1999) Proc NatlAcad Sci USA 96:8321-8323) or utilize such as sequence-specific recombinase or ribozyme (as follows) produces the target disappearance or sequence is reverse.

Also can realize the reduction of ε-cyclase quantity, function and/or activity by (for example by intermolecular homologous recombination method) in the sequence that nucleotide sequence (for example desire in the methods of the invention be inserted into nucleotide sequence) target is inserted into coding ε-cyclase.In the context of the present embodiment, preferably use contain at least a portion ε-cyclase gene sequence or flanking sequence and therefore can be in target cell specifically with the DNA construct of these sequences reorganization, thereby disappearance, add or substitute at least one nucleotides so that the functional reduction of ε-cyclase gene or the mode removed fully change ε-cyclase gene.This changes also can be at ε-cyclase gene controlling element (for example promoter), thereby coded sequence is remained unchanged, and stops or reducing and express (transcribe and/or translate).In conventional homologous recombination, 5 ' of the sequence of being inserted-and/or the 3 '-distolateral wing contain other have sufficient length and with ε-cyclase gene corresponding sequence (A and B) homology so that can produce the nucleotide sequence (A ' or B ') of homologous recombination.This length arrives (Thomas KR and Capecchi MR (1987) Cell 51:503 within several thousand base scopes in a hundreds of base usually; Strepp etc. (1998) Proc Natl Acad Sci USA95 (8): 4368-4373).For homologous recombination, use method described below to select the successfully clone of reorganization with the recombinant precursor transformed plant cells and based on resulting inactivation ε-cyclase.

In another preferred embodiment, by the raising recombination efficiency that combines with the method that promotes homologous recombination.Described these class methods, and comprised and for example express RecA protein, perhaps handled with the PARP inhibitor.Have been found that and utilize the PARP inhibitor can increase intrachromosomal homologous recombination in the tobacco plant ((1995) Plant J 7:203-210 such as Puchta H).By using these inhibitor, the efficient of homologous recombination rate and transgenic sequence disappearance can further improve in the special dna double chain fracture back recombinant precursor of induced sequence.Can use multiple PARP inhibitor.These inhibitor preferably include for example 3-aminobenzamide, 8-hydroxy-2-methyl quinazoline-4-one (NU1025), 1,11b-dihydro-[2H] chromene-[4,3,2-de] isoquinolin-3-ketone (GPI 6150), 5-aminoisoquinoline ketone, 3,4-dihydro-5-[4-(1-piperidyl) butoxy]-1-(2H) isoquinolines, or be described in material among WO 00/26192, WO 00/29384, WO 00/32579, WO 00/64878, WO 00/68206, WO 00/67734, WO 01/23386 and the WO 01/23390.

Other suitable method is that non-sense mutation is imported endogenous labelled protein plasmagene, for example by the RNA/DNA oligonucleotides being imported method (Zhu etc. (2000) the Nat Biotechnol18 (5): 555-558) of plant, perhaps knock out sudden change (Koncz etc. by producing such as T-DNA mutagenesis, PlantMol.Biol.1992,20 (5): 963-976).Just known " chimeric prosthetic (chimeraplasty) " also can produce point mutation (Cole-Strauss etc. (1999) NuclAcids Res 27 (5): 1323-1330 by the DNA-RNA heterozygote; Kmiec (1999) Gene therapy American Scientist87 (3): 240-247).

The dsRNAi method, by have a mind to the common inhibition of RNA and " VIGS " (" gene silencing of virus induction ") be also referred to as " PTGS " (PTGS) or " transcriptional gene silencing " (TGS).Since to the labelled protein plasmagene that will reduce and transgene expression intentionally or the demand of the homology between the dsRNA nucleotide sequence be lower than antisense method such as classics, so the PTGS/TGS method is particularly advantageous.Therefore, using labelled protein nucleotide sequence from species also can effectively reduce the expression of homology labelled protein in another species and not need separates and the structure explanation the labelled protein homologue that wherein exists.This has greatly reduced workload.

In the inventive method particularly preferred embodiment, compare with wild type and to reduce ε-cyclase activity by the following method:

A) with at least a double-stranded ε-cyclase RNA sequence guarantee an expression cassette of its expression or a plurality of expression cassette import plant and/or

B) with at least a ε-cyclase antisense RNA sequence or guarantee that the expression cassette of its expression imports plant.

In embodiment very particularly preferably, compare with wild type by with at least a double-stranded ε-cyclase RNA sequence or guarantee an expression cassette of its expression or a plurality of expression cassette imports plant and reduces ε-cyclase activity.

In preferred embodiments, use the genetically modified plant that in it is spent, has minimum ε-cyclase expression rate.

This by to spend special mode, particularly preferably is to reduce ε-cyclase activity in the special mode of floral leaf to realize preferably.

In above-mentioned particularly preferred embodiment, this is by spending under the specific promoter control or also being more preferably to transcribe ε-cyclase-dsRNA sequence under the control of floral leaf specific promoter to realize.

In another preferred embodiment, the plant of being cultivated is compared with wild type and also has at least a following activity rising that is selected from addition: the HMG-CoA reductase activity, (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity, 1-deoxidation-D-wood sugar-5-phosphoric acid-synthase activity, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity, prenyl diphosphate ester-Δ isomerase activity, Mang ox base-bisphosphate synthase activity, the farnesyl diphosphate synthase activity, Mang ox base Mang ox bisphosphate synthase activity, the phytoene synthase activity, the phytoene desaturase activity, sigma carotene desaturase activity, the crtISO activity, active and the MinD activity of FtsZ.

Think that the HMG-CoA reductase activity meaning is meant the HMG-CoA reductase (enzymatic activity of (3-hydroxy-3-methylglutaryl-coenzyme A reductase).

Think that the HMG-CoA reductase meaning is meant to have the protein that 3-hydroxy-3-methyl glutaryl coenzyme-A is converted into this enzymatic activity of mevalonic acid.

Therefore, think that the HMG-CoA reductase activity meaning is meant the amount of the 3-hydroxy-3-methyl glutaryl coenzyme-A that is transformed by protein HMG-CoA reductase in special time, or the amount of the mevalonic acid that forms.

Therefore, have the HMG-CoA reductase activity of comparing raising with wild type, promptly compare in special time the amount of the 3-hydroxy-3-methyl glutaryl coenzyme-A that transforms by protein HMG-CoA reductase or the amount of the mevalonic acid that forms increases with wild type.

Preferably, the HMG-CoA reductase activity is compared with wild type HMG-CoA reductase activity, improve at least 5%, more preferably improve at least 20%, more preferably improve at least 50%, more preferably improve at least 100%, more preferably improve at least 300%, even more preferably improve at least 500%, especially improve at least 600%.Think that the HMG-CoA reductase activity meaning is meant the enzymatic activity of HMG-CoA reductase.Genetically modified plant of the present invention and wild type or preferably measure under the following conditions with reference to HMG-CoA reductase activity in the plant:

In mortar, add liquid nitrogen and carry out extracting from 1: 1 to 1: 20 extraction buffer solution by the powerful grinding freezing vegetable material of homogenate and with ratio.Described ratio depends on obtainable enzymatic activity in the vegetable material, so that the mensuration of enzymatic activity and quantitatively can be in linear measures range.Generally, extract buffer solution and can contain 50mM HEPES-KOH (pH 7.4), 10mM MgCl ₂, 10mM KCl, 1mMEDTA, 1mM EGTA, 0.1% (v/v) Triton X-100,2mM EACA, 10% glycerine, 5mM KHCO ₃Before the extraction, add 2mM DTT and 0.5mM PMSF immediately.

Can measure HMG-CoA reductase activity (Schaller for example, Grausem, Benveniste, Chye, Tan, Song and Chua, Plant Physiol.109 (1995), 761-770 according to disclosed description; Chappell, Wolf, Proulx, Cuellar and Saunders, Plant Physiol.109 (1995) 1337-1343).Plant tissue can be at cold buffer solution (100mM potassium phosphate (pH 7.0), 4mM MgCl ₂, 5mM DTT) in carry out homogenate and extracting.With homogenate centrifugal 15 minutes of 4 ℃ of 10000g.Subsequently with supernatant once more with the centrifugal 45-60 of 100000g minute.Measure in the supernatant and HMG-CoA reductase activity in the microsomal fraction sediment (being resuspended in after 100mM potassium phosphate (pH 7.0) and the 50mM DTT).With every part of solution and suspension (protein content is equivalent to about 1-10 μ g in the suspension) in 30 ℃ contain 3mM NADPH and 20 μ M ( ¹⁴C) in the 100mM kaliumphosphate buffer (pH 7.0) of HMG-CoA (58 μ Ci/ μ M), volume of ideal is 26 μ l, hatches 15-60 minute.Add 5 μ l mevalonolactones (1mg/ml) and 6N HCl cessation reaction.After the adding, with mixture incubated at room 15 minutes.Formed in saturated potassium phosphate solution of 125 μ l (pH 6.0) and the 300 μ l ethyl acetate quantitative reactions by adding ( ¹⁴C) mevalonic acid.Mixture is fully mixed also centrifugal.By measuring the scintillation measuring radioactivity.

(E)-and 4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity, be also referred to as lytB or IspH, be considered to the enzymatic activity that the meaning is meant (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase.

Think that (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase meaning is meant to have the protein that (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate is converted into the enzymatic activity of prenyl diphosphate ester and dimethyl-allyl bisphosphate.

Therefore, think that (E)-4-hydroxy-3-methyl but-2-ene base-the bisphosphate reductase activity meaning is meant the amount of (the E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate that is transformed by protein (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase in special time, or the prenyl diphosphate ester that forms and the amount of dimethyl-allyl bisphosphate.

Therefore, when comparing (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity with wild type and improve, compare the amount of (the E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate that in special time, transforms with wild type, or the amount of prenyl diphosphate ester that forms and dimethyl-allyl bisphosphate increases by protein (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase.

Preferably, (E)-base-the bisphosphate reductase activity is compared increases by 5% at least for 4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity and wild type (E)-4-hydroxy-3-methyl but-2-ene, more preferably be at least 20%, more preferably be at least 50%, more preferably be at least 100%, more preferably be at least 300%, being more preferably increases by 500% at least, and especially at least 600%.

Genetically modified plant of the present invention and wild type or preferably measure under the following conditions with reference to (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity in the plant:

Can measure (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity by immunology detection.The generation of specific antibody is by Rohdich and the colleague (Rohdich that describes thereof, Hecht, G_rtner, Adam, Krieger, Amslinger, Arigoni, Bacher and Eisenreich: to the metabolism of the research of non-mevalonic acid terpenes biosynthesis pathway: IspH (LytB) albumen, Natl.Acad.Natl.Sci.USA 99 (2002), 1158-1163).Mensuration about catalytic activity, Altincicek and colleague (Altincicek thereof, Duin, Reichenberg, Hedderich, Kollas, Hintz, Wagner, Wiesner, Beck and Jomaa: in the isoprenoid biosynthesis, the final step of LytB albumen catalysis 2-C-methyl D-erythrite-4-phosphate approach; FEBS Letters 532 (2002) 437-440) describe a kind of vitro system tracking (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate and be reduced into prenyl diphosphate ester and dimethyl-allyl bisphosphate.

Think that the active meaning of 1-deoxidation-D-wood sugar-5-phosphate synthase is meant the enzymatic activity of 1-deoxidation-D-wood sugar-5-phosphate synthase.

Think that 1-deoxidation-D-wood sugar-5-phosphate synthase meaning is meant to have the protein that ethoxy-ThPP and glyceraldehyde 3-phosphate is converted into the enzymatic activity of 1-deoxidation-D-wood sugar-5-phosphoric acid.

Therefore, think that the active meaning of 1-deoxidation-D-wood sugar-5-phosphate synthase is meant the ethoxy-ThPP that transformed by protein 1-deoxidation-D-wood sugar-5-phosphate synthase and/or the amount of glyceraldehyde 3-phosphate in special time, or the amount of the 1-deoxidation-D-wood sugar-5-phosphoric acid that forms.

Therefore, when comparing the active raising of 1-deoxidation-D-wood sugar-5-phosphate synthase with wild type, compare the ethoxy-ThPP that in special time, transforms and/or the amount of glyceraldehyde 3-phosphate with wild type, or the amount of the 1-deoxidation-D-wood sugar-5-phosphoric acid that forms increases by protein 1-deoxidation-D-wood sugar-5-phosphate synthase.

Preferably, 1-deoxidation-D-wood sugar-5-phosphate synthase is active compares with wild type 1-deoxidation-D-wood sugar-5-phosphate synthase activity, improve at least 5%, more preferably improve at least 20%, more preferably improve at least 50%, more preferably improve at least 100%, more preferably improve at least 300%, even more preferably improve at least 500%, especially improve at least 600%.

In genetically modified plant of the present invention and wild type or in reference to plant, 1-deoxidation-D-wood sugar-5-phosphate synthase activity is preferably measured under the following conditions:

The reactant mixture (50-200 μ l) of measuring D-1-deoxy-D-xylulose sugar-5-phosphate synthase activity (DXS) contains 100mM Tris-HCl (pH 8.0), 3mM MgCl ₂, 3mM MnCl ₂, 3mM ATP, 1mM thiamine bisphosphate, 0.1% polysorbate60,1mM potassium fluoride, 30 μ M (2- ¹⁴C) pyruvic acid (0.5 μ Ci), 0.6mM DL-3-glyceraldehyde phosphate.Plant extracts was hatched in reaction solution 1-2 hour in 37 ℃.Subsequently, be heated to 80 ℃ of 3 minutes cessation reactions.Leave after the heart 5 minutes with per minute 13000, the evaporation supernatant is resuspended in 50 μ l methyl alcohol with residue, be applied to the TLC plate carry out thin-layer chromatography (silica gel 60, Merck is Darmstadt) and in normal propyl alcohol/ethyl acetate/water (6: 1: 3; V/v/v) separate in.Radiolabeled D-1-deoxy-D-xylulose sugar-5-phosphoric acid (or D-1-deoxy-D-xylulose sugar) and (2- ¹⁴C) pyruvic acid separates.Use scintillation counter to carry out quantitatively.This method has been described in Harker and Bramley (FEBS Letters 448 (1999) 115-119).In addition, the fluorimetry of mensuration DXS synthase activity is described (Analytical Biochemistry 296 (2001) 101-105) by Querol and colleague thereof.

Think that 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity meaning is meant the enzymatic activity of 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase.

Think that 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase meaning is meant to have 1-deoxidation-D-wood sugar-5-phosphoric acid is converted into the protein of the enzymatic activity of beta carotene.

Therefore, think that 1-deoxidation-D-wood sugar-the 5-phosphoric acid reduction isomerase activity meaning is meant the amount of the 1-deoxidation-D-wood sugar-5-phosphoric acid that is transformed by protein 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase in special time, or the amount of the prenyl diphosphate ester that forms.

Therefore, when comparing 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity with wild type and improve, compare the amount of the 1-deoxidation-D-wood sugar-5-phosphoric acid that in special time, transforms with wild type, or the amount of the prenyl diphosphate ester that forms increases by protein 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase.

Preferably, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity is compared with wild type 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity, improve at least 5%, more preferably improve at least 20%, more preferably improve at least 50%, more preferably improve at least 100%, more preferably improve at least 300%, even more preferably improve at least 500%, especially improve at least 600%.

In genetically modified plant of the present invention and wild type or in reference to plant, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity is preferably measured under the following conditions:

The activity of D-1-deoxy-D-xylulose sugar-5-phosphoric acid reduction isomerase (DXR) is containing 100mMTris-HCl (pH 7.5), 1mM MnCl ₂, 0.3mM NADPH and 0.3mM 1-deoxy-D-xylulose-4-phosphoric acid buffer solution in measure, wherein 1-deoxy-D-xylulose-4-phosphoric acid can zymetology synthesize, (Kuzuyama for example, Takahashi, Watanabe and Seto:Tetrahedon letters39 (1998) 4509-4512).Add plant extracts and begin reaction.Reaction volume is generally 0.2 to 0.5ml; Hatched 30-60 minute for 37 ℃.Between incubation period, use the oxidation of photometer tracking and measuring NADPH at the 340nm place.

Think that prenyl diphosphate ester Δ-isomerase activity meaning is meant the enzymatic activity of isopentene group-bisphosphate Δ-isomerase.

Think that isopentene group-bisphosphate Δ-isomerase meaning is meant to have the protein that the prenyl diphosphate ester is converted into the enzymatic activity of dimethyl-allyl phosphate.

Therefore, think that isopentene group-bisphosphate Δ-isomerase activity meaning is meant the amount of the prenyl diphosphate ester that is transformed by isopentene group-bisphosphate Δ-isomerase in special time or the amount of the dimethyl-allyl phosphate that forms.

Therefore, when comparing isopentene group-bisphosphate Δ-isomerase activity with wild type and improve, compare the amount of the prenyl diphosphate ester that in special time, transforms with wild type or the amount of the dimethyl-allyl phosphate that forms increases by protein isopentene group-bisphosphate Δ-isomerase.

Preferably, isopentene group-bisphosphate Δ-isomerase activity is compared with wild type isopentene group-bisphosphate Δ-isomerase activity, improve at least 5%, more preferably improve at least 20%, more preferably improve at least 50%, more preferably improve at least 100%, more preferably improve at least 300%, even more preferably improve at least 500%, especially improve at least 600%.

In genetically modified plant of the present invention and wild type or in reference to plant, isopentene group-bisphosphate Δ-isomerase activity is preferably measured under the following conditions:

According to can measure the active (Fraser of isopentene group-bisphosphate isomerase (IPP isomerase) by the Fraser and the disclosed method of working together thereof, R_mer, Shipton, Mills, Kiano, Misawa, Drake, Schuch and Bramley: the transgene tomato plant of extra phytoene synthase is expressed in assessment in the fruit specific mode; Proc.Natl.Acad.Sci.USA 99 (2002), 1092-1097, and it is based on Fraser, Pinto, Holloway and Bramley, Plant Journal 24 (2000), 551-558).For enzymatic determination, use 0.5 μ Ci (1- ¹⁴C) IPP (isopentenyl pyrophosphate ester) (56mCi/mmol, Amersham plc) is containing 1mM DTT, 4mM MgCl as substrate ₂, 6mM MnCl ₂, 3mM ATP, 0.1% polysorbate60,1mM potassium fluoride 0.4M Tris-HCl (pH 8.0) in volume be that about 150-500 μ l is hatched.Extract is mixed (for example with 1: 1 ratio) with buffer solution and hatched at least 5 hours at 28 ℃.Subsequently, add about 200 μ l methyl alcohol and hatch at 37 ℃ and carried out acid hydrolysis in about 1 hour by adding concentrated hydrochloric acid (final concentration is 25%).Then, use benzinum (mixing) to carry out twice and repeat extracting (each 500 μ l) with 10% diethyl ether.The use scintillation counter is measured the radioactivity in each upper strata phase (hyperphase).The short incubation period of using 5 minutes can be measured the specific enzymes activity because the formation of short reaction time inhibitory reaction accessory substance (see L ü tzow and Beyer: isopentene group in the daffodil chromoplast-bisphosphate Δ-isomerase and with the relation of phytoene synthase complex; Biochim.Biophys.Acta 959 (1988), 118-126).

Think that Mang ox base-bisphosphate synthase activity meaning is meant the enzymatic activity of Mang ox base-bisphosphate synthase.

Think that the Mang ox base-bisphosphate synthase meaning is meant to have the protein that prenyl diphosphate ester and dimethyl-allyl phosphate is converted into the enzymatic activity of Mang ox base-bisphosphate.

Therefore, think that the Mang ox base-bisphosphate synthase activity meaning is meant the prenyl diphosphate ester that transformed by protein Mang ox base-bisphosphate synthase and/or the amount of dimethyl-allyl phosphate in special time, or the amount of the Mang ox base-bisphosphate that forms.

Therefore, when comparing Mang ox base-bisphosphate synthase activity with wild type and improve, compare the prenyl diphosphate ester that in special time, transforms and/or the amount of dimethyl-allyl phosphate with wild type, or the amount of the Mang ox base-bisphosphate that forms increases by protein Mang ox base-bisphosphate synthase.

Preferably, Mang ox base-bisphosphate synthase activity is compared with wild type Mang ox base-bisphosphate synthase activity, improve at least 5%, more preferably improve at least 20%, more preferably improve at least 50%, more preferably improve at least 100%, more preferably improve at least 300%, even more preferably improve at least 500%, especially improve at least 600%.

In genetically modified plant of the present invention and wild type or in reference to plant, Mang ox base-bisphosphate synthase activity is preferably measured under the following conditions:

Can be determined at 50mM Tris-HCl (pH 7.6), 10mMMgCl after adding plant extracts ₂, 5mM MnCl ₂, 2mM DTT, 1mM ATP, 0.2% Tween-20,5 μ M ( ¹⁴C) activity of the Mang ox base-bisphosphate synthase (GPP synthase) among IPP and the 50 μ M DMAPP (dimethylallylpyrophosphate ester) is (according to Bouvier, Suire, d ' Harlingue, Backhaus and Camara: the molecular cloning of Mang ox base-bisphosphate synthase and the compartmentation that monoterpene synthesizes in plant cell, Plant Journal 24 (2000) 241-252).Hatch for 37 ℃, after 2 hours, with the product dephosphorylation (according to Koyama, Fuji and Ogura: the enzymatic hydrolysis of polyisoprene pyrophosphate, Methods Enzymol.110 (1985), 153-155) also mix determination method analysis (Dogbo with TLC and radioactivity, Bardat, Quennemet and Camara: the metabolism of plastid terpenoid: synthetic by the external inhibition phytoene of phenethyl pyrophosphate (phenethyl pyrophosphate) derivative, FEBS Letters 219 (1987) 211-215).

Think that the farnesyl diphosphate synthase activity meaning is meant the enzymatic activity of farnesyl diphosphate synthase.

Think that the farnesyl diphosphate synthase meaning is meant to have the protein that Mang ox base bisphosphate and prenyl diphosphate ester is converted into the enzymatic activity of farnesyl diphosphate.

Therefore, the farnesyl diphosphate synthase activity is the Mang ox base bisphosphate that transformed by protein farnesyl bisphosphate synthase in special time and the amount of prenyl diphosphate ester, or the amount of the farnesyl diphosphate that forms.

Therefore, when comparing the raising of farnesyl diphosphate synthase activity with wild type, compare the amount of basic bisphosphate of the Mang ox that in special time, transforms and prenyl diphosphate ester with wild type, or the amount of the farnesyl diphosphate that forms increases by protein farnesyl bisphosphate synthase.

Preferably, the farnesyl diphosphate synthase activity is compared with wild type farnesyl diphosphate synthase activity, improve at least 5%, more preferably improve at least 20%, more preferably improve at least 50%, more preferably improve at least 100%, more preferably improve at least 300%, even more preferably improve at least 500%, especially improve at least 600%.

In genetically modified plant of the present invention and wild type or in reference to plant, the farnesyl diphosphate synthase activity is preferably measured under the following conditions:

According to the method for Joly and Edwards can measure farnesylpyrophosphate synthase (FPP synthase) activity (Journal of Biological Chemistry 268 (1993), 26983-26989).In view of the above, at 10mM HEPES (pH 7.2), 1mM MgCl ₂, 1mM dithiothreitol (DTT), 20 μ M Mang oxen base pyrophosphate and 40 μ M (1- ¹⁴C) measure enzymatic activity in isopentenyl pyrophosphate ester (4Ci/mmol) buffer solution.Reactant mixture is hatched at 37 ℃; Add 2.5N HCl (in 70% ethanol that contains 19 μ g/ml farnesol) cessation reaction.Then by carrying out acid hydrolysis hydrolysis product in 30 minutes at 37 ℃.Add 10% NaOH neutralise mixt, and carry out extracting by vibrating with hexane.Utilizing scintillation counter to measure every part of hexane mixes to measure radioactivity.

In addition, after plant extracts and radiolabeled IPP hatched, (Silica-Gel SE60 Merck) can reaction product isolated by thin-layer chromatography in benzene/methyl alcohol (9: 1).The radiolabeled product of wash-out and measure radioactivity (according to Gaffe, Bru, Causse, Vidal, Stamitti-Bert, Carde and Gallusci: a kind of tomato farnesylpyrophosphate gene LEFPS1 of high expressed during the fruit early development; Plant Physiology 123 (2000) 1351-1362).

Think that the Mang ox base Mang ox base bisphosphate synthase activity meaning is meant the enzymatic activity of Mang ox base Mang ox base bisphosphate synthase.

Think that the Mang ox base Mang ox base bisphosphate synthase meaning is meant to have the protein that farnesyl diphosphate and prenyl diphosphate ester is converted into the enzymatic activity of Mang ox base Mang ox base bisphosphate.

Therefore, think that the Mang ox base Mang ox base bisphosphate synthase activity meaning is meant the farnesyl diphosphate that transformed by protein Mang ox base Mang ox base bisphosphate synthase and/or the amount of prenyl diphosphate ester in special time, or the amount of the Mang ox that forms base Mang ox base bisphosphate.

Therefore, when comparing the raising of Mang ox base Mang ox base bisphosphate synthase activity with wild type, compare the farnesyl diphosphate that in special time, transforms and/or the amount of prenyl diphosphate ester with wild type, or the amount of the Mang ox that forms base Mang ox base bisphosphate increases by protein Mang ox base Mang ox base bisphosphate synthase.

Preferably, Mang ox base Mang ox base bisphosphate synthase activity is compared with wild-type activity, improve at least 5%, more preferably improve at least 20%, more preferably improve at least 50%, more preferably improve at least 100%, more preferably improve at least 300%, even more preferably improve at least 500%, especially improve at least 600%.

In genetically modified plant of the present invention and wild type or in reference to plant, Mang ox base Mang ox base bisphosphate synthase activity is preferably measured under the following conditions:

Can measure the activity (Biochim.Biophys.Acta 920 (1987), 140-148: by affinity chromatography from capsicum chromoplast purifying isopentenyl pyrophosphate ester isomerase and Mang ox base Mang ox base pyrophosphate synthase) of Mang ox base Mang ox base pyrophosphate synthase (GGPP synthase) according to Dogbo and the described method of Camara.Therefore, plant extracts is joined buffer solution (50mM Tris-HCl (pH 7.6), the 2mM MgCl that cumulative volume is approximately 200 μ l ₂, 1mM MnCl ₂, 2mM dithiothreitol (DTT), (1- ¹⁴C) IPP (0.1 μ Ci, 10 μ M), 15 μ M DMAPP, GPP or FPP) in.Hatch 1-2 hour (or longer) at 30 ℃.Add 0.5ml ethanol and 0.1ml 6N HCl cessation reaction.37 ℃ hatched 10 minutes after, with 6N NaOH neutralization reaction mixture, mixes with 1ml water and vibrates and carry out extracting with the 4ml ether.Utilize scintillation counter to measure radioactivity in a ether phase (as 0.2ml).In addition, after the acid hydrolysis, but by the radiolabeled prenol of vibration extracting in ether and by HPLC (25cm Spherisorb ODS-1 post, 5 μ m; With methanol (90: 10; V/v) with 1ml/ minute flow velocity wash-out) separate, and by monitoring radioactivity quantitative (Wiedemann, Misawa and Sandmann :) from the purifying and the enzymatic property of Mang ox base Mang ox base pyrophosphate synthase behind expression in escherichia coli of having a liking for summer spore Erwinia (Erwiniauredovora).

Think that the active meaning of phytoene synthase is meant the enzymatic activity of phytoene synthase.

Think that the phytoene synthase meaning is meant to have the protein that the terminal linear residue of lycopene is converted into the enzymatic activity of β-ionone ring.

Particularly, think that the phytoene synthase meaning is meant to have the protein that Mang ox base Mang ox base bisphosphate is converted into the enzymatic activity of phytoene.

Therefore, think that the active meaning of phytoene synthase is meant the amount of the Mang ox base Mang ox base bisphosphate that is transformed by the protein phytoene synthase in special time, or the amount of the phytoene that forms.

Therefore, when comparing the active raising of phytoene synthase with wild type, compare the amount of the Mang ox base Mang ox base bisphosphate that in special time, transforms with wild type by the protein phytoene synthase, or the increase of the amount of the phytoene that forms.

Preferably, think that phytoene synthase is active and compare with wild type phytoene synthase activity, improve at least 5%, more preferably improve at least 20%, more preferably improve at least 50%, more preferably improve at least 100%, more preferably improve at least 300%, even more preferably improve at least 500%, especially improve at least 600%.

In genetically modified plant of the present invention and wild type or in reference to plant, the phytoene synthase activity is preferably measured under the following conditions:

Can measure the active (Fraser of phytoene synthase (PSY) by the Fraser and the disclosed method of working together thereof, Romer, Shipton, Mills, Kiano, Misawa, Drake, Schuch and Bramley: the transgene tomato plant is expressed the assessment of extra phytoene synthase in the fruit specific mode, and Proc.Natl.Acad.Sci.USA 99 (2002), 1092-1097, it is based on Fraser, Pinto, Holloway and Bramley, Plant Journal 24 (2000) 551-558).For enzyme assay, with ( ³H) (15mCi/mM, AmericanRadiolabeled Chemicals St.Louis) are containing 1mM DTT, 4mMMgCl as substrate to Mang ox base Mang ox base pyrophosphate ₂, 6mM MnCl ₂, 3mM ATP, 0.1% polysorbate60,1mM potassium fluoride 0.4MTris-HCl (pH 8.0) in carry out.Plant extracts is mixed with buffer solution, is that 500 μ l are hatched with 295 μ l buffer solutions and extract with cumulative volume for example.Hatched at least 5 hours at 28 ℃.Then, with twice (each 500 μ l) vibration of chloroform extracting phytoene.In course of reaction, use methanol (95: 5; V/v) on silica plate, utilize TLC to separate formed radiolabeled phytoene.(by heating several iodine crystal) can identify phytoene on silica plate under rich iodine environment.With the phytoene standard items as reference.Amount by scintillation counter determination method detection of radioactive labels product.In addition, utilize also quantitative phytoene (Fraser, the Albrecht and Sandmann: develop the precursor of high performance liquid chromatography system of HPLC that radioactivity seeker is housed to be used for separating radiolabeled carrotene and forming in specific enzymatic reaction; J.Chromatogr.645 (1993) 265-272).

Think that the active meaning of phytoene desaturase is meant the enzymatic activity of phytoene desaturase.

Think that the phytoene desaturase meaning is meant to have the protein that phytoene is converted into phytofluene and/or phytofluene is converted into the enzymatic activity of sigma carotene (zeta-carrotene).

Therefore, think that the active meaning of phytoene desaturase is meant phytoene or the amount of phytofluene or the amount of phytofluene that forms or sigma carotene that is transformed by the protein phytoene desaturase in special time.

Therefore, when comparing the active raising of phytoene desaturase with wild type, compare phytoene or the amount of phytofluene or the amount increase of phytofluene that forms or sigma carotene that in special time, transforms with wild type by the protein phytoene desaturase.

Preferably, think that phytoene desaturase is active and compare with wild type phytoene desaturase activity, improve at least 5%, more preferably improve at least 20%, more preferably improve at least 50%, more preferably improve at least 100%, more preferably improve at least 300%, even more preferably improve at least 500%, especially improve at least 600%.

In genetically modified plant of the present invention and wild type or in reference to plant, the phytoene desaturase activity is preferably measured under the following conditions:

By with radiolabeled ( ¹⁴C) phytoene mixes unsaturated carrotene and can measure the activity of phytoene desaturase (PDS) (as R_mer, Fraser, Kiano, Shipton, Misawa, Schuch and Bramley report: provitamin A content in the assessment transgene tomato plant; Nature Biotechnology 18 (2000) 666-669).Can be as the phytoene (Fraser of synthesizing radioactive mark as described in the Fraser, De la Rivas, Mackenzie, Bramley: phycomyces blakesleeanus (Phycomyces blakesleanus) CarB mutant: their purposes in measuring phytoene desaturase; Phytochemistry 30 (1991), 3971-3976).Can with contain 10mM MgCl ₂In the cumulative volume of 1ml, hatch the plastid film of target tissue with the 100mM MES buffer solution (pH 6.0) of 1mM dithiothreitol (DTT).Add be dissolved in the acetone ( ¹⁴C) phytoene (each situation hatch next time about 100000 the fission/minute), wherein acetone concentration is no more than 5% (v/v).In the dark mixture was hatched about 6-7 hour 28 ℃ of vibrations.Subsequently, separate also quantitatively with three extracting pigments of about 5ml benzinum (mixing) and with the HPLC method with 10% ether.

In addition, can such as Fraser etc. report measure the activity (Fraser of phytoene desaturase, Misawa, Linden, Yamano, Kobayashi and Sandmann: expression in escherichia coli, purifying and reactivation reorganization have a liking for summer spore Erwinia phytoene desaturase, Journal ofBiological Chemistry 267 (1992), 19891-19895).

Think that the active meaning of sigma carotene desaturase is meant the enzymatic activity of sigma carotene desaturase.

Think that the sigma carotene desaturase meaning is meant to have the protein that sigma carotene is converted into neurosporene and/or neurosporene is converted into the enzymatic activity of lycopene.

Therefore, think that the active meaning of sigma carotene desaturase is meant the sigma carotene that transformed by protein sigma carotene desaturase or the amount of neurosporene in special time, or the neurosporene that forms or the amount of lycopene.

Therefore, when comparing the active raising of sigma carotene desaturase with wild type, compare the sigma carotene that in special time, transforms or the amount of neurosporene with wild type, or the amount of neurosporene that forms or lycopene increases by protein sigma carotene desaturase.

Preferably, think that the sigma carotene desaturase is active and compare with wild type sigma carotene desaturase activity, improve at least 5%, more preferably improve at least 20%, more preferably improve at least 50%, more preferably improve at least 100%, more preferably improve at least 300%, even more preferably improve at least 500%, especially improve at least 600%.

In genetically modified plant of the present invention and wild type or in reference to plant, sigma carotene desaturase activity is preferably measured under the following conditions:

Can in 0.2M potassium phosphate (pH 7.8, the about 1ml of buffer solution volume), measure the analysis of sigma carotene desaturase (ZDS desaturase).Breitenbach and colleague thereof disclose the analytic approach of measuring, and (Takaichi and Sandmann: the higher plant type of expression and purifying is from the catalysis characteristics of the sigma carotene desaturase of capsicum for Breitenbach, Kuntz; European Journal ofBiochemistry.265 (1): 376-383, in October, 1999).Every kind of assay determination mixture contains 3mg and is suspended in phosphatid ylcholine, 5 μ g sigma carotenes or neurosporene, 0.02% butylated hydroxytoluene, 10 μ l decyl plastoquinones (1mM methyl alcohol is stored liquid) and plant extracts in the 0.4M kaliumphosphate buffer (pH 7.8).The volume of plant extracts is adjusted to the active amount that exists of ZDS desaturase so that may carry out quantitatively in linear measurement range.Generally when being in about 28 ℃, dark hatched about 17 hours by thermal agitation (200 rev/mins).By adding 4ml acetone 10 minutes extracting carotenoid of vibration 50 ℃ the time.In this mixture, carotenoid is transferred to benzinum phase (containing 10% ether).Under nitrogen, evaporation ether/benzinum phase is dissolved in 20 μ l again with carotenoid and utilizes the HPLC method to separate with quantitative.

Think that the active meaning of crtlSO is meant the enzymatic activity of crtlSO albumen.

Think that the crtlSO albumen meaning is meant to have 7,9,7 ', 9 '-four-cis-lycopene is converted into the protein of the enzymatic activity of alltrans-lycopene.

Therefore, think that the active meaning of crtlSO is meant in special time by 7,9 of protein b-cyclase conversion, the amount of the amount of 7 ', 9 '-four-cis-lycopene or the alltrans-lycopene of formation.

Therefore, when comparing with wild type that crtlSO is active to be improved, compare with wild type in special time by crtlSO albumen transform 7,9, the amount of 7 ', 9 '-four-cis-lycopene, or the amount of the alltrans-lycopene that forms increases.

Preferably, crtlSO is active to compare with wild type crtlSO activity, improve at least 5%, more preferably improve at least 20%, more preferably improve at least 50%, more preferably improve at least 100%, more preferably improve at least 300%, even more preferably improve at least 500%, especially improve at least 600%.

In genetically modified plant of the present invention and wild type or in reference to plant, the crtlSO activity is preferably measured under the following conditions:

Think that the active meaning of FtsZ is meant the physiologically active of FtsZ albumen.

Think the FtsZ albumen meaning be meant have promote cell division and plastid division is active and with the protein of microtubular protein homology.

Think that the active meaning of MinD is meant the physiologically active of MinD protein.

Think that the MinD protein meaning is meant the protein that has multi-function action in cell division.It is the relevant ATP enzyme of a kind of film and can demonstrates oscillating movement from a utmost point to another utmost point in cell.

And the activity increase of the enzyme of non-mevalonate pathway can cause the further increase of purpose keto-acid carotenoid (ketocarotenoid) end-product.The example is 4-cytidine diphosphate (CDP) (diphosphocytidyl)-2-C-methyl D-erythrite synthase; 4-cytidine diphosphate (CDP)-2-C-methyl D-erythrite kinases and 2-C-methyl D-erythrite-2; 4-ring bisphosphate synthase.By modifying the gene expression of corresponding gene, can improve the activity of described enzyme.Change by the concentration that antibody and corresponding engram technology can detect the respective egg white matter with standard mode.HMG-CoA/ ( E )-4--3--2--/1--D--5-/1--D--5-/-Δ-/-/////ζ-/crtlSO/FtsZ/MinD； For example by expressing and protein level is closed restricted regulatory mechanism, or the gene expression of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid by increasing coding HMG-CoA reductase and/or coding ( E )-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase and/or coding 1-deoxidation-D-wood sugar-5-phosphate synthase and/or coding 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase and/or coding isopentene group-bisphosphate Δ-isomerase and/or coding Mang ox base-bisphosphate synthase and/or coding farnesyl diphosphate synthase and/or coding Mang ox base Mang ox base bisphosphate synthase and/or coding phytoene synthase and/or coding phytoene desaturase and/or coding sigma carotene desaturase and/or coding crtlSO albumen and/or the nucleic acid of coding FtsZ albumen and/or the MinD protein of encoding.

The gene expression of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of the nucleic acid of coding HMG-CoA reductase and/or coding ( E )-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase and/or coding 1-deoxidation-D-wood sugar-5-phosphate synthase and/or coding 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase and/or coding isopentene group-bisphosphate Δ-isomerase and/or coding Mang ox base-bisphosphate synthase and/or coding farnesyl diphosphate synthase and/or coding Mang ox base Mang ox base bisphosphate synthase and/or coding phytoene synthase and/or coding phytoene desaturase and/or coding sigma carotene desaturase and/or coding crtlSO albumen and/or coding FtsZ albumen and/or coding MinD protein is compared with wild type to be improved; For example induce HMG-CoA reductase gene and/or ( E )-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase gene and/or 1-deoxidation-D-wood sugar-5-phosphate synthase gene and/or 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase gene and/or isopentene group-bisphosphate Δ-isomerase gene and/or Mang ox base-bisphosphate synthase gene and/or farnesyl diphosphate synthase gene and/or Mang ox base Mang ox base bisphosphate synthase gene and/or phytoene synthase gene and/or phytoene desaturase gene and/or sigma carotene delta 8 desaturase genes and/or crtlSO gene and/or Fts gene and/or MinD gene by activating son; HMG-CoA/ ( E )-4--3--2--/1--D--5-/1--D--5-/-Δ-/-/////ζ-/crtlSO/Fts/MinD,HMG-CoA/ ( E )-4--3--2--/1--D--5-/1--D--5-/-Δ-/-/////ζ-/crtlSO/FtsZ/MinD。

HMG-CoA/ ( E )-4--3--2--/1--D--5-/1--D--5-/-Δ-/-/////ζ-/crtlSO/FtsZ/MinD、HMG-CoA/ ( E )-4--3--2--/1--D--5-/1--D--5-/-Δ-/-/////ζ-/crtlSO/FtsZ/MinD。

For example, this can be achieved by modifying corresponding promoter DNA sequence.For example disappearance or the insertion by dna sequence dna can realize that this kind causes the modification that the gene expression rate improves.

In preferred embodiments; HMG-CoA/ ( E )-4--3--2--/1--D--5-/1--D--5-/-Δ-/-/////ζ-/crtlSO/FtsZ/MinD,HMG-CoA/ ( E )-4--3--2--/1--D--5-/1--D--5-/-Δ-/-/////ζ-/crtlSO/FtsZ/MinD。

Therefore; Basically, can use arbitrary HMG-CoA reductase gene or (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase gene or 1-deoxidation-D-wood sugar-5-phosphate synthase gene or 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase gene or isopentene group-bisphosphate Δ-isomerase gene or Mang ox base-bisphosphate synthase gene or farnesyl diphosphate synthase gene or Mang ox base Mang ox base bisphosphate synthase gene or phytoene synthase gene or phytoene desaturase gene or sigma carotene delta 8 desaturase genes or crtlSO gene or FtsZ gene or MinD gene.

When host plant can not be expressed the respective egg white matter, in the time of maybe can not being endowed the ability of expressing the respective egg white matter, in the genome HMG-CoA reductase sequence that contains introne in eucaryon source or (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase sequence or 1-deoxidation-D-wood sugar-5-phosphate synthase sequence or 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase sequence or isopentene group-bisphosphate Δ-isomerase sequence or Mang ox base-bisphosphate synthase sequence or farnesyl diphosphate synthase sequence or Mang ox base Mang ox base bisphosphate synthase sequence or phytoene synthase sequence or phytoene desaturase sequence or sigma carotene desaturase sequence or crtlSO sequence or FtsZ sequence or MinD sequence in, preferably use through manufactured nucleotide sequence in advance, as corresponding cDNA.

Therefore, in this embodiment preferred, compare with wild type, in the preferred genetically modified plants of the present invention, contain at least one extra HMG-CoA reductase gene and/or (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase gene and/or 1-deoxidation-D-wood sugar-5-phosphate synthase gene and/or 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase gene and/or isopentene group-bisphosphate Δ-isomerase gene and/or Mang ox base-bisphosphate synthase gene and/or farnesyl diphosphate synthase gene and/or Mang ox base Mang ox base bisphosphate synthase gene and/or phytoene synthase gene and/or phytoene desaturase gene and/or sigma carotene delta 8 desaturase genes and/or crtlSO gene and/or FtsZ gene and/or MinD gene.

In this preferred embodiment; Genetically modified plant has; ,HMG-CoAHMG-CoA/ ( E )-4--3--2-- ( E )-4--3--2--/1--D--5-1--D--5-/1--D--5-1--D--5-/-Δ--Δ-/--/////ζ-ζ-/crtlSOcrtlSO/FtsZFtsZ/MinDMinD。

The example of HMG-CoA reductase gene is:

Accession number be NM 106299 the nucleic acid from the coding HMG-CoA reductase of mouseearcress (Arabidopsis thaliana) (nucleic acid: SEQ ID NO:111, protein: SEQ ID NO:112),

And comprise from other organic other HMG-CoA reductase genes with following accession number:

P54961，P54870，P54868，P54869，O02734，P22791，P54873，P54871，P23228，P13704，P54872，Q01581，P17425，P54874，P54839，P14891，P34135，O64966，P29057，P48019，P48020，P12683，P43256，Q9XEL8，P34136，O64967，P29058，P48022，Q41437，P12684，Q00583，Q9XHL5，Q41438，Q9YAS4，O76819，O28538，Q9Y7D2，P54960，O51628，P48021，Q03163，P00347，P14773，Q12577，Q59468，P04035，O24594，P09610，Q58116，O26662，Q01237，Q01559，Q12649，O74164，O59469，P51639，Q10283，O08424，P20715，P13703，P13702，Q96UG4，Q8SQZ9，O15888，Q9TUM4，P93514，Q39628，P93081，P93080，Q944T9，Q40148，Q84MM0，Q84LS3，Q9Z9N4，Q9KLM0

(E)-example of 4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase gene is:

From the nucleic acid (lytB/ISPH) of mouseearcress coding (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase, accession number AY168881, (nucleic acid: SEQ ID NO:113, protein: SEQ ID NO:114),

And comprise from other organic other (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase genes with following accession number:

T04781，AF270978_1，NP_485028.1，NP_442089.1，NP_681832.1，ZP_00110421.1，ZP_00071594.1，ZP_00114706.1，ISPH_SYNY3，ZP_00114087.1，ZP_00104269.1，AF398145_1，AF398146_1，AAD55762.1，AF514843_1，NP_622970.1，NP_348471.1，NP_562001.1，NP_223698.1，NP_781941.1，ZP_00080042.1，NP_859669.1，NP_214191.1，ZP_00086191.1，ISPH_VIBCH，NP_230334.1，NP_742768.1，NP_302306.1，ISPH_MYCLE，NP_602581.1，ZP_00026966.1，NP_520563.1，NP_253247.1，NP_282047.1，ZP_00038210.1，ZP_00064913.1，CAA61555.1，ZP_00125365.1，ISPH_ACICA，EAA24703.1，ZP_00013067.1，ZP_00029164.1，NP_790656.1，NP_217899.1，NP_641592.1，NP_636532.1，NP_719076.1，NP_660497.1，NP_422155.1，NP_715446.1，ZP_00090692.1，NP_759496.1，ISPH_BURPS，ZP_00129657.1，NP_215626.1，NP_335584.1，ZP_00135016.1，NP_789585.1，NP_787770.1，NP_769647.1，ZP_00043336.1，NP_242248.1，ZP_00008555.1，NP_246603.1，ZP_00030951.1，NP_670994.1，NP_404120.1，NP_540376.1，NP_733653.1，NP_697503.1，NP_840730.1，NP_274828.1，NP_796916.1，ZP_00123390.1，NP_824386.1，NP_737689.1，ZP_00021222.1，NP_757521.1，NP_390395.1，ZP_00133322.1，CAD76178.1，NP_600249.1，NP_454660.1，NP_712601.1，NP_385018.1，NP_751989.1

The example of 1-deoxidation-D-wood sugar-5-phosphate synthase gene is:

From the nucleic acid of the coding 1-deoxidation-D-wood sugar-5-phosphate synthase that eats tomato, accession number #AF143812 (nucleic acid: SEQ ID NO:115, protein: SEQ ID NO:116),

And comprise from other organic other 1-deoxidations-D-wood sugar-5-phosphate synthase genes with following accession number:

AF143812_1，DXS_CAPAN，CAD22530.1，AF182286_1，NP_193291.1，T52289，AAC49368.1，AAP14353.1，D71420，DXS_ORYSA，AF443590_1，BAB02345.1，CAA09804.2，NP_850620.1，CAD22155.2，AAM65798.1，NP_566686.1，CAD22531.1，AAC33513.1，CAC08458.1，AAG10432.1，T08140，AAP14354.1，AF428463_1，ZP_00010537.1，NP_769291.1，AAK59424.1，NP_107784.1，NP_697464.1，NP_540415.1，NP_196699.1，NP_384986.1，ZP_00096461.1，ZP_00013656.1，NP_353769.1，BAA83576.1，ZP_00005919.1，ZP_00006273.1，NP_420871.1，AAM48660.1，DXS_RHOCA，ZP_00045608.1，ZP_00031686.1，NP_841218.1，ZP_00022174.1，ZP_00086851.1，NP_742690.1，NP?520342.1，ZP_00082120.1，NP_790545.1，ZP_00125266.1，CAC17468.1，NP_252733.1，ZP_00092466.1，NP_439591.1，NP_414954.1，NP_752465.1，NP_622918.1，NP_286162.1，NP_836085.1，NP_706308.1，ZP_00081148.1，NP_797065.1，NP_213598.1，NP_245469.1，ZP_00075029.1，NP_455016.1，NP_230536.1，NP_459417.1，NP_274863.1，NP_283402.1，NP_759318.1，NP_406652.1，DXS_SYNLE，DXS_SYNP7，NP_440409.1，ZP_00067331.1，ZP_00122853.1，NP_717142.1，ZP_00104889.1，NP_243645.1，NP_681412.1，DXS_SYNEL，NP_637787.1，DXS_CHLTE，ZP_00129863.1，NP_661241.1，DXS_XANCP，NP_470738.1，NP_484643.1，ZP_00108360.1，NP_833890.1，NP_846629.1，NP_658213.1，NP_642879.1，ZP_00039479.1，ZP_00060584.1，ZP_00041364.1，ZP_00117779.1，NP_299528.1

The example of 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase gene is:

From the nucleic acid of coding 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase of mouseearcress, accession number #AF148852, (nucleic acid: SEQ ID NO:137, protein: SEQ ID NO:138),

And comprise from other organic other 1-deoxidations-D-wood sugar-5-phosphoric acid reduction isomerase genes with following accession number:

AF148852，AY084775，AY054682，AY050802，AY045634，AY081453，AY091405，AY098952，AJ242588，AB009053，AY202991，NP_201085.1，T52570，AF331705_1，BAB16915.1，AF367205_1，AF250235_1，CAC03581.1，CAD22156.1，AF182287_1，DXR_MENPI，ZP_00071219.1，NP_488391.1，ZP_00111307.1，DXR_SYNLE，AAP56260.1，NP_681831.1，NP_442113.1，ZP_00115071.1，ZP_00105106.1，ZP_00113484.1，NP_833540.1，NP_657789.1，NP_661031.1，DXR_BACHD，NP_833080.1，NP_845693.1，NP_562610.1，NP_623020.1，NP_810915.1，NP_243287.1，ZP_00118743.1，NP_464842.1，NP_470690.1，ZP_00082201.1，NP_781898.1，ZP_00123667.1，NP_348420.1，NP_604221.1，ZP_00053349.1，ZP_00064941.1，NP_246927.1，NP_389537.1，ZP_00102576.1，NP_519531.1，AF124757_19，DXR_ZYMMO，NP_713472.1，NP_459225.1，NP_454827.1，ZP_00045738.1，NP_743754.1，DXR_PSEPK，ZP_00130352.1，NP_702530.1，NP_841744.1，NP_438967.1，AF514841_1，NP_706118.1，ZP_00125845.1，NP_404661.1，NP_285867.1，NP_240064.1，NP_414715.1，ZP_00094058.1，NP_791365.1，ZP_00012448.1，ZP_00015132.1，ZP_00091545.1，NP_629822.1，NP_771495.1，NP_798691.1，NP_231885.1，NP_252340.1，ZP_00022353.1，NP_355549.1，NP_420724.1，ZP_00085169.1，EAA17616.1，NP_273242.1，NP_219574.1，NP_387094.1，NP_296721.1，ZP_00004209.1，NP_823739.1，NP_282934.1，BAA77848.1，NP_660577.1，NP_760741.1，NP_641750.1，NP_636741.1，NP_829309.1，NP_298338.1，NP_444964.1，NP_717246.1，NP_224545.1，ZP_00038451.1，DXR_KITGR，NP_778563.1.

The example of isopentene group-bisphosphate Δ-isomerase gene is:

Carry from John Burroughs and to receive the nucleic acid (ipiAa1) of coding isopentene group-bisphosphate Δ-isomerase of adonis amurensis clone ApIPI28, accession number #AF188060, by Cunningham, F.X.Jr. and Gantt, E.: by the multigene family of the isopentene group-bisphosphate isomerase of encoding in the complementary plant identification of the allos in Escherichia coli, Plant Cell Physiol.41 (1), the open (nucleic acid: SEQ ID NO:117 of 119-123 (2000), protein: SEQ ID NO:118)

And comprise from other organic other isopentene groups-bisphosphate Δ-isomerase genes with following accession number:

Q38929，O48964，Q39472，Q13907，O35586，P58044，O42641，O35760，Q10132，P15496，Q9YB30，Q8YNH4，Q42553，O27997，P50740，O51627，O48965，Q8KFR5，Q39471，Q39664，Q9RVE2，Q01335，Q9HHE4，Q9BXS1，Q9KWF6，Q9CIF5，Q88WB6，Q92BX2，Q8Y7A5，Q8TT35Q9KK75，Q8NN99，Q8XD58，Q8FE75，Q46822，Q9HP40，P72002，P26173，Q9Z5D3，Q8Z3X9，Q8ZM82，Q9X7Q6，O13504，Q9HFW8，Q8NJL9，Q9UUQ1，Q9NH02，Q9M6K9，Q9M6K5，Q9FXR6，O81691，Q9S7C4，Q8S3L8，Q9M592，Q9M6K3，Q9M6K7，Q9FV48，Q9LLB6，Q9AVJ1，Q9AVG8，Q9M6K6，Q9AVJ5，Q9M6K2，Q9AYS5，Q9M6K8，Q9AVG7，Q8S3L7，Q8W250，Q94IE1，Q9AVI8，Q9AYS6，Q9SAY0，Q9M6K4，Q8GVZ0，Q84RZ8，Q8KZ12，Q8KZ66，Q8FND7，Q88QC9，Q8BFZ6，BAC26382，CAD94476.

The example of Mang ox base-bisphosphate synthase gene is:

From the nucleic acid of coding Mang ox base-bisphosphate synthase of mouseearcress, accession number #Y17376, Bouvier, F., Suire, C., d ' Harlingue, A., Backhaus, R.A. and Camara, B.: the molecular cloning of Mang ox base-bisphosphate synthase and the compartmentation that monoterpene synthesizes in plant cell, Plant is (2) J.24,241-252 (2000) (nucleic acid: SEQ ID NO:119, protein: SEQ ID NO:120)

And comprise from other organic other Mang ox base-bisphosphate synthase genes with following accession number:

Q9FT89，Q8LKJ2，Q9FSW8，Q8LKJ3，Q9SBR3，Q9SBR4，Q9FET8，Q8LKJ1，Q84LG1，Q9JK86

The example of farnesyl diphosphate synthase gene is:

Accession number #U80605 from the nucleic acid (FPS1) of the coding farnesyl diphosphate synthase of mouseearcress by Cunillera, N., Arro, M., Delourme, D., Karst, F., Boronat, A. and Ferrer, A.: mouseearcress comprises the farnesyl diphosphate synthase gene that two species diversity are expressed, J.Biol.Chem.271 (13), 7774-7780 (1996) is open, (nucleic acid: SEQ ID NO:121, protein: SEQ ID NO:122)

And comprise from other organic other farnesyl diphosphate synthase genes with following accession number:

P53799，P37268，Q02769，Q09152，P49351，O24241，Q43315，P49352，O24242，P49350，P08836，P14324，P49349，P08524，O66952，Q08291，P54383，Q45220，P57537，Q8K9A0，P22939，P45204，O66126，P55539，Q9SWH9，Q9AVI7，Q9FRX2，Q9AYS7，Q94IE8，Q9FXR9，Q9ZWF6，Q9FXR8，Q9AR37，O50009，Q94IE9，Q8RVK7，Q8RVQ7，O04882，Q93RA8，Q93RB0，Q93RB4，Q93RB5，Q93RB3，Q93RB1，Q93RB2，Q920E5.

The example of Mang ox base Mang ox base bisphosphate synthase gene is:

From the nucleic acid of the basic Mang ox base of the coding Mang ox of sinapsis alba (Sinapis alba) bisphosphate synthase, accession number #X98795 is by Bonk, M., Hoffmann, B., Von Lintig, J., Schledz, M., Al-Babili, S., Hobeika, E., Kleinig, H. and Beyer, P.: the chromoplast input of external four kinds of carotenoid biosynthetic enzymes has disclosed the different fate before film combination and oligomerization assembling, Eur.J.Biochem.247 (3), the open (nucleic acid: SEQ ID NO:123 of 942-950 (1997), protein: SEQ ID NO:124)

And comprise from other organic other Mang ox base Mang ox base bisphosphate synthase genes with following accession number:

P22873，P34802，P56966，P80042，Q42698，Q92236，O95749，Q9WTN0，Q50727，P24322，P39464，Q9FXR3，Q9AYN2，Q9FXR2，Q9AVG6，Q9FRW4，Q9SXZ5，Q9AVJ7，Q9AYN1，Q9AVJ4，Q9FXR7，Q8LSC5，Q9AVJ6，Q8LSC4，Q9AVJ3，Q9SSU0，Q9SXZ6，Q9SST9，Q9AVJ0，Q9AVI9，Q9FRW3，Q9FXR5，Q94IF0，Q9FRX1，Q9K567，Q93RA9，Q93QX8，CAD95619，EAA31459

The example of phytoene synthase gene is:

From the nucleic acid of the coding phytoene synthase of Erwinia uredovora, accession number #D90087 is by Misawa, N., Nakagawa, M., Kobayashi, K., Yamano, S., Izawa, Y., Nakamura, K. and Harashima, K.: set forth the carotenoid biosynthesis pathway of Erwinia uredovora by the gene outcome of expression in escherichia coli being carried out functional analysis; J.Bacteriol.172 (12), 6704-6712 (1990) is open, (nucleic acid: SEQ ID NO:125, protein: SEQ ID NO:126),

And comprise from other organic other phytoene synthase genes with following accession number:

CAB39693，BAC69364，AAF10440，CAA45350，BAA20384，AAM72615，BAC09112，CAA48922，P_001091，CAB84588，AAF41518，CAA48155，AAD38051，AAF33237，AAG10427，AAA34187，BAB73532，CAC19567，AAM62787，CAA55391，AAB65697，AAM45379，CAC27383，AAA32836，AAK07735，BAA84763，P_000205，AAB60314，P_001163，P_000718，AAB71428，AAA34153，AAK07734，CAA42969，CAD76176，CAA68575，P_000130，P_001142，CAA47625，CAA85775，BAC14416，CAA79957，BAC76563，P_000242，P_000551，AAL02001，AAK15621，CAB94795，AAA91951，P_000448

The example of phytoene desaturase gene is:

From the nucleic acid of the coding phytoene desaturase of Erwinia uredovora, accession number #D90087 is by Misawa, N., Nakagawa, M., Kobayashi, K., Yamano, S., Izawa, Y., Nakamura, K. and Harashima, K.: set forth the carotenoid biosynthesis pathway of Erwinia uredovora by the gene outcome of expression in escherichia coli being carried out functional analysis; J.Bacteriol.172 (12), 6704-6712 (1990) is open, (nucleic acid: SEQ ID NO:127, protein: SEQ IDNO:128),

And comprise from other organic other phytoene desaturase genes with following accession number:

AAL15300，A39597，CAA42573，AAK51545，BAB08179，CAA48195，BAB82461，AAK92625，CAA55392，AAG10426，AAD02489，AAO24235，AAC12846，AAA99519，AAL38046，CAA60479，CAA75094，ZP_001041，ZP_001163，CAA39004，CAA44452，ZP_001142，ZP_000718，BAB82462，AAM45380，CAB56040，ZP_001091，BAC09113，AAP79175，AAL80005，AAM72642，AAM72043，ZP_000745，ZP_001141，BAC07889，CAD55814，ZP_001041，CAD27442，CAE00192，ZP_001163，ZP_000197，BAA18400，AAG10425，ZP_001119，AAF13698，2121278A，AAB35386，AAD02462，BAB68552，CAC85667，AAK51557，CAA12062，AAG51402，AAM63349，AAF85796，BAB74081，AAA91161，CAB56041，AAC48983，AAG14399，CAB65434，BAB73487，ZP_001117，ZP_000448，CAB39695，CAD76175，BAC69363，BAA17934，ZP_000171，AAF65586，ZP_000748，BAC07074，ZP_001133，CAA64853，BAB74484，ZP_?001156，AAF23289，AAG28703，AAP09348，AAM71569，BAB69140，ZP_000130，AAF41516，AAG18866，CAD95940，NP_656310，AAG10645，ZP_000276，ZP_000192，ZP_000186，AAM94364，EAA31371，ZP_000612，BAC75676，AAF65582

The example of sigma carotene delta 8 desaturase genes is:

Nucleic acid from the coding sigma carotene desaturase of daffodil, accession number #AJ224683, by Al-Babili, S., Oelschlegel, J. and Beyer, P.: from the cDNA (accession number AJ224683) of the coding beta carotene desaturase of daffodil (PGR98-103), Plant Physiol.117, the open (nucleic acid: SEQ ID NO:129 of 719-719 (1998), protein: SEQ ID NO:130)

And comprise from other organic other sigma carotene delta 8 desaturase genes with following accession number:

Q9R6X4，Q38893，Q9SMJ3，Q9SE20，Q9ZTP4，O49901，P74306，Q9FV46，Q9RCT2，ZDS_NARPS，BAB68552.1，CAC85667.1，AF372617_1，ZDS_TARER，CAD55814.1，CAD27442.1，2121278A，ZDS_CAPAN，ZDS_LYCES，NP_187138.1，AAM63349.1，ZDS_ARATH，AAA91161.1，ZDS_MAIZE，AAG14399.1，NP_441720.1，NP_486422.1，ZP_00111920.1，CAB56041.1，ZP_00074512.1，ZP_00116357.1，NP_681127.1，ZP_00114185.1，ZP_00104126.1，CAB65434.1，NP_662300.1

The example of crtlSO gene is:

Nucleic acid from edible tomato coding crtlSO; Accession number #AF416727, by Isaacson, T., Ronen, G., Zamir, D.and Hirschberg, J.: cloning orange element (tangerine) from tomato, to be disclosed in the plant carotenoid isomerase be essential for producing beta carotene and lutein; Plant Cell 14 (2), 333-342 (2002) open (nucleic acid: SEQ ID NO:131, protein: SEQ ID NO:132),

And comprise from other organic other crtlSO gene: AAM53952 with following accession number

The example of FtsZ gene is:

From the nucleic acid of the coding FtsZ of marigold, accession number #AF251346, by Moehs, C.P., Tian, L., Osteryoung, K.W. and Dellapenna, D.: the analysis of between the pot marigold petal puberty, the carotenoid biosynthesis gene being expressed; Plant Mol.Biol.45 (3), 281-293 (2001) open (nucleic acid: SEQ ID NO:133, protein: SEQ ID NO:134),

And comprise from other organic other FtsZ genes with following accession number:

CAB89286.1，AF205858_1，NP_200339.1，CAB89287.1，CAB41987.1，AAA82068.1，T06774，AF383876_1，BAC57986.1，CAD22047.1，BAB91150.1，ZP_00072546.1，NP_440816.1，T51092，NP_683172.1，BAA85116.1，NP_487898.1，JC4289，BAA82871.1，NP_781763.1，BAC57987.1，ZP_00111461.1，T51088，NP_190843.1，ZP_00060035.1，NP_846285.1，AAL07180.1，NP_243424.1，NP_833626.1，AAN04561.1，AAN04557.1，CAD22048.1，T51089，NP_692394.1，NP_623237.1，NP_565839.1，T51090，CAA07676.1，NP_113397.1，T51087，CAC44257.1，E84778，ZP_00105267.1，BAA82091.1，ZP_00112790.1，BAA96782.1，NP_348319.1，NP_471472.1，ZP_00115870.1，NP_465556.1，NP_389412.1，BAA82090.1，NP_562681.1，AAM22891.1，NP_371710.1，NP_764416.1，CAB95028.1，FTSZ_STRGR，AF120117_1，NP_827300.1，JE0282，NP_626341.1，AAC45639.1，NP_785689.1，NP_336679.1，NP_738660.1，ZP_00057764.1，AAC32265.1，NP_814733.1，FTSZ_MYCKA，NP_216666.1，CAA75616.1，NP_301700.1，NP_601357.1，ZP_00046269.1，CAA70158.1，ZP_00037834.1，NP_268026.1，FTSZ_ENTHR，NP_787643.1，NP_346105.1，AAC32264.1，JC5548，AAC95440.1，NP_710793.1，NP_687509.1，NP_269594.1，AAC32266.1，NP_720988.1，NP_657875.1，ZP_00094865.1，ZP_00080499.1，ZP_00043589.1，JC7087，NP_660559.1，AAC46069.1，AF179611_14，AAC44223.1，NP_404201.1.

The example of MinD gene is:

From the nucleic acid of the coding MinD of marigold, accession number #AF251019, by Moehs, C.P., Tian, L., Osteryoung, K.W. and Dellapenna, D.: the analysis of between the pot marigold petal puberty, the carotenoid biosynthesis gene being expressed; Plant Mol.Biol.45 (3), 281-293 (2001) open (nucleic acid: SEQ ID NO:135, protein: SEQ ID NO:136),

And comprise from other organic other MinD genes with following accession number:

NP_197790.1，BAA90628.1，NP_038435.1，NP_045875.1，AAN33031.1，NP_050910.1，CAB53105.1，NP_050687.1，NP_682807.1，NP_487496.1，ZP_00111708.1，ZP_00071109.1，NP_442592.1，NP_603083.1，NP_782631.1，ZP_00097367.1，ZP_00104319.1，NP_294476.1，NP_622555.1，NP_563054.1，NP_347881.1，ZP_00113908.1，NP_834154.1，NP_658480.1，ZP_00059858.1，NP_470915.1，NP_243893.1，NP_465069.1，ZP_00116155.1，NP_390677.1，NP_692970.1，NP_298610.1，NP_207129.1，ZP_00038874.1，NP_778791.1，NP_223033.1，NP_641561.1，NP_636499.1，ZP_00088714.1，NP_213595.1，NP_743889.1，NP_231594.1，ZP_00085067.1，NP_797252.1，ZP_00136593.1，NP_251934.1，NP_405629.1，NP_759144.1，ZP_00102939.1，NP_793645.1，NP_699517.1，NP_460771.1，NP_860754.1，NP_456322.1，NP_718163.1，NP_229666.1，NP_357356.1，NP_541904.1，NP_287414.1，NP_660660.1，ZP_00128273.1，NP_103411.1，NP_785789.1，NP_715361.1，AF149810_1，NP_841854.1，NP_437893.1，ZP_00022726.1，EAA24844.1，ZP_00029547.1，NP_521484.1，NP_240148.1，NP_770852.1，AF345908_2，NP_777923.1，ZP_00048879.1，NP_579340.1，NP_143455.1，NP_126254.1，NP_142573.1，NP_613505.1，NP_127112.1，NP_712786.1，NP_578214.1，NP_069530.1，NP_247526.1，AAA85593.1，NP_212403.1，NP_782258.1，ZP_00058694.1，NP_247137.1，NP_219149.1，NP_276946.1，NP_614522.1，ZP_00019288.1，CAD78330.1

In above-mentioned embodiment preferred, as the HMG-CoA reductase gene, the preferred nucleic acid that uses contains amino acid sequence SEQ ID NO:112 as coding or by this sequence being carried out amino acid replacement, insertion or lacking the protein of institute's calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ ID NO:112 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of HMG-CoA reductase.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of corresponding reverse translation and the homology between the SEQ ID NO:112 from database, can be easily from find other example of HMG-CoA reductase and HMG-CoA reductase gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:111 initial by hybridization and round pcr in a manner known way also can be easily from the multiple organism discovery HMG-CoA reductase of its genome sequence the unknown and other example of HMG-CoA reductase gene.

In particularly preferred embodiment more, in order to increase the activity of HMG-CoA reductase, such nucleic acid is imported organism, the protein of described nucleic acid coding contains the amino acid sequence of the HMG-CoA reductase of sequence SEQ IDNO:112.

For example, suitable nucleotide sequence can obtain by the reverse translation peptide sequence according to genetic code.

Therefore, preferably use those codons of selecting frequent use according to the special codon of plant.Be easy to determine the codon selection based on computer evaluation to relevant other known of organism.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:111.

In above-mentioned embodiment preferred, as (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase gene, the preferred nucleic acid coding that uses contains amino acid sequence SEQ ID NO:114 or by this sequence is carried out amino acid replacement, insert or lack the protein of institute's calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ ID NO:114 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase.

As mentioned above, by relatively from the amino acid sequence of database or the nucleotide sequence and the homology between the SEQ ID NO:114 of corresponding reverse translation, can be easily from find such as the known multiple organism of its genome sequence (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase with (E)-other example of 4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase gene.

In addition, as mentioned above, such as by sequence SEQ ID NO:113 initial by hybridization and round pcr in a manner known way also can be easily from the multiple organism of its genome sequence the unknown find (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase with (E)-other example of 4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase gene.

In particularly preferred embodiment more, in order to increase the activity of (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase, such nucleic acid is imported organism, and the protein of described nucleic acid coding contains the amino acid sequence of (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase of sequence SEQ ID NO:114

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:113.

In above-mentioned embodiment preferred, as 1-deoxidation-D-wood sugar-5-phosphate synthase gene, the preferred nucleic acid that uses contains amino acid sequence SEQ ID NO:116 as coding or by this sequence is carried out amino acid replacement, insert or lack the protein of institute's calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ ID NO:116 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of 1-deoxidation-D-wood sugar-5-phosphate synthase.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of corresponding reverse translation and the homology between the SEQ ID NO:116 from database, can be easily from find other example of 1-deoxidation-D-wood sugar-5-phosphate synthase and 1-deoxidation-D-wood sugar-5-phosphate synthase gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:115 initial by hybridization and round pcr in a manner known way also can be easily from multiple organism discovery 1-deoxidation-D-wood sugar-5-phosphate synthase of its genome sequence the unknown and other example of 1-deoxidation-D-wood sugar-5-phosphate synthase gene.

In particularly preferred embodiment more, in order to increase the activity of 1-deoxidation-D-wood sugar-5-phosphate synthase, such nucleic acid is imported organism, and the protein of described nucleic acid coding contains the amino acid sequence of 1-deoxidation-D-wood sugar-5-phosphate synthase of sequence SEQID NO:116.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:115.

In above-mentioned embodiment preferred, as 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase gene, the preferred nucleic acid that uses contains amino acid sequence SEQ ID NO:138 as coding or by this sequence is carried out amino acid replacement, insert or lack the protein of institute's calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ ID NO:138 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of corresponding reverse translation and the homology between the SEQ ID NO:138 from database, can be easily from find other example of 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase and 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:137 initial by hybridization and round pcr in a manner known way also can be easily from multiple organism discovery 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase of its genome sequence the unknown and other example of 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase gene.

In particularly preferred embodiment more, in order to increase the activity of 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase, such nucleic acid is imported organism, and the protein of described nucleic acid coding contains the amino acid sequence of sequence SEQ ID NO:138 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:137.

In above-mentioned embodiment preferred, as isopentene group D-isomerase gene, the preferred nucleic acid coding that uses contains amino acid sequence SEQ ID NO:118 or by this sequence being carried out the protein of amino acid replacement, insertion or disappearance institute calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQID NO:118 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of isopentene group D-isomerase.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of corresponding reverse translation and the homology between the SEQ ID NO:118 from database, can be easily from find other example of isopentene group D-isomerase and isopentene group D-isomerase gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:117 initial by hybridization and round pcr in a manner known way also can be easily from the multiple organism discovery isopentene group D-isomerase of its genome sequence the unknown and other example of isopentene group D-isomerase gene.

In particularly preferred embodiment more, in order to increase the activity of isopentene group D-isomerase, such nucleic acid is imported organism, the protein of described nucleic acid coding contains the amino acid sequence of sequence SEQ IDNO:118 isopentene group D-isomerase.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:117.

In above-mentioned embodiment preferred, as Mang ox base-bisphosphate synthase gene, the preferred nucleic acid coding that uses contains amino acid sequence SEQ ID NO:120 or by this sequence being carried out the protein of amino acid replacement, insertion or disappearance institute calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ ID NO:120 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of Mang ox base-bisphosphate synthase.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of corresponding reverse translation and the homology between the SEQ ID NO:120 from database, can be easily from find other example of Mang ox base-bisphosphate synthase and Mang ox base-bisphosphate synthase gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:119 initial by hybridization and round pcr in a manner known way also can be easily from multiple organism discovery Mang ox base-bisphosphate synthase of its genome sequence the unknown and other example of Mang ox base-bisphosphate synthase gene.

In particularly preferred embodiment more, in order to increase the activity of Mang ox base-bisphosphate synthase, such nucleic acid is imported organism, and described nucleic acid coding contains the protein of the amino acid sequence of sequence SEQ ID NO:120 Mang ox base-bisphosphate synthase.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:119.

In above-mentioned embodiment preferred, as the farnesyl diphosphate synthase gene, the preferred nucleic acid coding that uses contains amino acid sequence SEQ ID NO:122 or by this sequence being carried out the protein of amino acid replacement, insertion or disappearance institute calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ ID NO:122 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of farnesyl diphosphate synthase.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of corresponding reverse translation and the homology between the SEQ ID NO:122 from database, can be easily from find other example of farnesyl diphosphate synthase and farnesyl diphosphate synthase gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:121 initial by hybridization and round pcr in a manner known way also can be easily from the multiple organism discovery farnesyl diphosphate synthase of its genome sequence the unknown and other example of farnesyl diphosphate synthase gene.

In particularly preferred embodiment more, in order to increase the activity of farnesyl diphosphate synthase, such nucleic acid is imported organism, described nucleic acid coding contains the protein of the amino acid sequence of sequence SEQ ID NO:122 farnesyl diphosphate synthase.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:121.

In above-mentioned embodiment preferred, as Mang ox base Mang ox base bisphosphate synthase gene, the preferred nucleic acid coding that uses contains amino acid sequence SEQ ID NO:124 or by this sequence is carried out amino acid replacement, insert or lack the protein of institute's calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ ID NO:124 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of Mang ox base Mang ox base bisphosphate synthase.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of corresponding reverse translation and the homology between the SEQ ID NO:124 from database, can be easily from find other example of Mang ox base Mang ox base bisphosphate synthase and the basic Mang ox base of Mang ox bisphosphate synthase gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as in a manner known way also can be easily finding other example of Mang ox base Mang ox base bisphosphate synthase and the basic bisphosphate synthase gene of Mang ox base Mang ox by hybridization and round pcr from the multiple organism of its genome sequence the unknown by sequence SEQ ID NO:123 is initial.

In particularly preferred embodiment more, in order to increase the activity that Mang ox base Mang ox base bisphosphate closes, such nucleic acid is imported organism, and described nucleic acid coding contains the protein of the amino acid sequence that sequence SEQ IDNO:124 Mang ox base Mang ox base bisphosphate closes.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:123.

In above-mentioned embodiment preferred, as the phytoene synthase gene, the preferred nucleic acid coding that uses contains amino acid sequence SEQ ID NO:126 or by this sequence being carried out the protein of amino acid replacement, insertion or disappearance institute calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQID NO:126 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of phytoene synthase.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of corresponding reverse translation and the homology between the SEQ ID NO:126 from database, can be easily from find other embodiment of phytoene synthase and phytoene synthase gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:125 initial by hybridization and round pcr in a manner known way also can be easily from the multiple organism discovery phytoene synthase of its genome sequence the unknown and other example of phytoene synthase gene.

In particularly preferred embodiment more, in order to increase the activity of phytoene synthase, such nucleic acid is imported organism, described nucleic acid coding contains the protein of the amino acid sequence of sequence SEQ ID NO:126 phytoene synthase.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:125.

In above-mentioned embodiment preferred, as the phytoene desaturase gene, the preferred nucleic acid coding that uses contains amino acid sequence SEQ ID NO:128 or by this sequence being carried out the protein of amino acid replacement, insertion or disappearance institute calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ ID NO:128 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of phytoene desaturase.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of corresponding reverse translation and the homology between the SEQ ID NO:128 from database, can be easily from find other example of phytoene desaturase and phytoene desaturase gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:127 initial by hybridization and round pcr in a manner known way also can be easily from the multiple organism discovery phytoene desaturase of its genome sequence the unknown and other example of phytoene desaturase gene.

In particularly preferred embodiment more, in order to increase the activity of phytoene desaturase, such nucleic acid is imported organism, and described nucleic acid coding contains the protein of the amino acid sequence of sequence SEQ ID NO:128 phytoene desaturase.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:127.

In above-mentioned embodiment preferred, as the sigma carotene delta 8 desaturase genes, the preferred nucleic acid coding that uses contains amino acid sequence SEQ ID NO:130 or by this sequence being carried out the protein of amino acid replacement, insertion or disappearance institute calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ ID NO:130 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of sigma carotene desaturase.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of corresponding reverse translation and the homology between the SEQ ID NO:130 from database, can be easily from find other example of sigma carotene desaturase and sigma carotene delta 8 desaturase genes such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:129 initial by hybridization and round pcr in a manner known way also can be easily from the multiple organism discovery sigma carotene desaturase of its genome sequence the unknown and other example of sigma carotene delta 8 desaturase genes.

In particularly preferred embodiment more, in order to increase the activity of sigma carotene desaturase, such nucleic acid is imported organism, described nucleic acid coding contains the protein of the amino acid sequence of sequence SEQ ID NO:130 sigma carotene desaturase.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:129.

In above-mentioned embodiment preferred, as the Crtlso gene, the preferred nucleic acid coding that uses contains amino acid sequence SEQ ID NO:132 or by this sequence being carried out the protein of amino acid replacement, insertion or disappearance institute calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ IDNO:132 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of Crtlso.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of respective opposed translation and the homology between the SEQ ID NO:132 from database, can be easily from find other example of Crtlso and Crtlso gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:131 initial by hybridization and round pcr in a manner known way also can be easily from the multiple organism discovery Crtlso of its genome sequence the unknown and other example of Crtlso gene.

In particularly preferred embodiment more, in order to increase the activity of Crtlso, such nucleic acid is imported organism, described nucleic acid coding contains the protein of the amino acid sequence of sequence SEQ ID NO:132 Crtlso.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:131.

In above-mentioned embodiment preferred, as the FtsZ gene, the preferred nucleic acid coding that uses contains amino acid sequence SEQ ID NO:134 or by this sequence being carried out the protein of amino acid replacement, insertion or disappearance institute calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ ID NO:134 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of FtsZ.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of respective opposed translation and the homology between the SEQ ID NO:134 from database, can be easily from find other example of FtsZ and FtsZ gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:133 initial by hybridization and round pcr in a manner known way also can be easily from the multiple organism discovery FtsZ of its genome sequence the unknown and other example of FtsZ gene.

In particularly preferred embodiment more, in order to increase the activity of FtsZ, such nucleic acid is imported organism, described nucleic acid coding contains the protein of the amino acid sequence of sequence SEQ ID NO:134 FtsZ.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:133.

In above-mentioned embodiment preferred, as the MinD gene, the preferred nuclear coding that uses contains amino acid sequence SEQ ID NO:136 or by this sequence being carried out the protein of amino acid replacement, insertion or disappearance institute calling sequence, at amino acid levels, the homogeneity of aftermentioned protein and sequence SEQ ID NO:136 is at least 30%, be preferably at least 50%, more preferably be at least 70%, even more preferably be at least 90%, be most preferably at least 95%, and have the enzymatic property of MinD.

As mentioned above, by comparing amino acid sequence or the nucleotide sequence of respective opposed translation and the homology between the SEQ ID NO:136 from database, can be easily from find other example of MinD and MinD gene such as the known multiple organism of its genome sequence.

In addition, as mentioned above, such as by sequence SEQ ID NO:135 initial by hybridization and round pcr in a manner known way also can be easily from the multiple organism discovery MinD of its genome sequence the unknown and other embodiment of MinD gene.

In particularly preferred embodiment more, in order to increase the activity of MinD, such nucleic acid is imported organism, described nucleic acid coding contains the protein of the amino acid sequence of sequence SEQ ID NO:136 MinD.

The nucleic acid importing organism that will contain in particularly preferred embodiments, sequence SEQ ID NO:135.

In addition, pass through in a manner known way from the chemical synthesis of nucleotide structure unit, for example, can prepare all above-mentioned HMG-CoA reductase genes by the fragment condensation of each overlapping complementary nucleic acid construction unit of double helix, (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase gene, 1-deoxidation-D-wood sugar-5-phosphate synthase gene, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase gene, isopentene group-bisphosphate Δ-isomerase gene, Mang ox base-bisphosphate synthase gene, method acyl-bisphosphate synthase gene, Mang ox base Mang ox base bisphosphate synthase gene, the phytoene synthase gene, the phytoene desaturase gene, the sigma carotene delta 8 desaturase genes, the crtlSO gene, FtsZ gene or MinD gene.For example, can carry out the chemical synthesis (Voet, Voet, second edition, Wiley Press New York, 896-897 page or leaf) of oligonucleotides in known manner by phosphoramidite method.The annealing of synthetic oligonucleotide, the Klenow fragment fill up the gap that utilizes archaeal dna polymerase, coupled reaction and conventional cloning process are described in (1989) such as Sambrook, Molecularcloning:A laboratory manual, publishing house of cold spring harbor laboratory.

In the embodiment that method is more preferably, plant additionally has compares the endogenous B-hydroxylase activity that has reduced with wild type.

As mentioned above, think the activity that has reduced preferably be meant based on different cell biology mechanism parts suppress fully basically or blocking-up plant cell, plant or its part, tissue, organ, cell or seed in the function of enzyme.

For example, by the amount of protein or the amount of mRNA in the minimizing plant, can realize that the activity of comparing in the plant with wild type reduces.Therefore, compare the activity that has reduced with wild type and can directly measure, or compare the amount of protein in the plant of the present invention or the amount of mRNA with wild type and measure by measuring.

Active reduction comprise protein quantity be reduced to lack fully basically this protein (that is to say shortage can detected corresponding activity or lack immunology can detected respective egg white matter).

Think that the active meaning of endogenous B-hydroxylase is meant the enzymatic activity of the B-hydroxylase that endogenous plant is intrinsic.

Think that the endogenous B-hydroxylase meaning is meant the hydroxylase that endogenous as mentioned above plant is intrinsic.If for example marigold is the target plant that carries out genetic modification, think that the endogenous B-hydroxylase is meant the B-hydroxylase of marigold.

Therefore think that the endogenous B-hydroxylase meaning specifically is meant to have the intrinsic protein of plant that beta carotene is converted into this enzymatic activity of luteole.

Therefore, think that the active meaning of endogenous B-hydroxylase is meant the amount of the beta carotene that is transformed by protein endogenous B-hydroxylase in special time or the amount of the luteole that forms.

When comparing with wild type that the endogenous B-hydroxylase is active to be reduced, compare the amount of the beta carotene that in special time, transforms with wild type or the amount of the luteole that forms reduces by protein endogenous B-hydroxylase.

Preferably, the endogenous B-hydroxylase is active to reduce at least 5%, more preferably for reducing at least 20%, more preferably for reducing at least 50%, more preferably for reducing by 100%.Particularly preferably, endogenous B-hydroxylase activity is closed fully.

Be surprisingly found out that producing for example plant of lutein for example in the plants of tagetes species of carotenoid in a large number that by the alpha-carotene approach it is active and to increase the allos hydroxylase activity as required be favourable to reduce the endogenous B-hydroxylase.Particularly preferably, the hydroxylase that uses or its function equivalent are from the plant that produces a large amount of carotenoid by the beta carotene approach, for example be above-mentioned B-hydroxylase (nucleic acid: SEQ ID No.107, protein: SEQ ID No.108) from tomato.

Measure endogenous B-hydroxylase activity in the mode similar as mentioned above to measuring hydroxylase activity.

Preferably, reduce endogenous B-hydroxylase activity in the plant by at least a following method:

A) import at least a double-stranded endogenous B-hydroxylase RNA sequence, hereinafter be also referred to as endogenous B-hydroxylase-dsRNA, or import an expression cassette or a plurality of expression cassette of guaranteeing its expression.

Comprise wherein endogenous B-hydroxylase-dsRNA at those methods of endogenous Beta-hydroxylase gene (just referring to genomic dna sequence) or endogenous B-hydroxylase transcript (just referring to the mRNA sequence) such as promoter sequence,

B) import at least a endogenous B-hydroxylase antisense RNA sequence, hereinafter be also referred to as endogenous B-hydroxylase-antisense RNA, or import the expression cassette of guaranteeing its expression.Comprise that endogenous B-hydroxylase-antisense RNA wherein is at endogenous Beta-hydroxylase gene (just referring to genomic dna sequence) or endogenous Beta-hydroxylase gene transcript (just referring to the RNA sequence)

Those methods.Also comprise α-different nucleotide sequence.

C) expression cassette that imports at least a endogenous B-hydroxylase-antisense RNA that combines with ribozyme or guarantee its expression.

D) import at least a endogenous B-hydroxylase and have a mind to RNA sequence, hereinafter be also referred to as endogenous B-hydroxylase-have a mind to RNA inducing common inhibition, or import the expression cassette of guaranteeing its expression

E) import at least a at endogenous Beta-hydroxylase gene, B-hydroxylase RNA or B-hydroxylase protein DNA-or protein-binding factor or guarantee the expression cassette of its expression

F) import at least a nucleic acid sequence or guarantee the expression cassette of its expression, cause the degraded of endogenous B-hydroxylase RNA

G) import at least one construct in the endogenous Beta-hydroxylase gene, to produce afunction, for example produce the displacement in terminator codon or the reading frame, for example insert by in the endogenous Beta-hydroxylase gene, producing, disappearance, oppositely or sudden change implement.Preferably, target inserts described endogenous Beta-hydroxylase gene or implanting needle and can produce the method for the sequence-specific nuclease of endogenous Beta-hydroxylase gene sequence and knock out sudden change by homologous recombination.

The known additive method of those skilled in the art also can be used among the present invention to reduce endogenous B-hydroxylase or its activity or function.For example import the dominant variant of endogenous B-hydroxylase or guarantee that the expression cassette of its expression also can be favourable.Every kind of diverse ways all can cause the amount of endogenous B-hydroxylase protein, amount and/or active reduction of mRNA.Also can consider to be used in combination these methods.Additive method also is known for those skilled in the art and can comprises inhibition or stop the transhipment of the processing of endogenous B-hydroxylase, luteole epoxidase and/or endogenous B-hydroxylase or its mRNA, extension or the termination that the inhibition ribosomes adheres to, suppresses the RNA montage, induces endogenous B-hydroxylase-RNA-digestive enzyme and/or suppress to translate.

Can each preferable methods be described by exemplary embodiment hereinafter:

A) import double-stranded endogenous B-hydroxylase RNA sequence (endogenous B-hydroxylase-dsRNA)

Deeply described above be used to reduce ε-cyclase activity carry out the method for gene regulation in the double-stranded RNA mode.In a similar manner, can implement this method to reduce endogenous B-hydroxylase activity.

Think that double-stranded endogenous B-hydroxylase RNA sequence or endogenous B-hydroxylase-dsRNA preferably are meant to have zone that contains duplex structure and the RNA molecule that contains following nucleotide sequence in this zone:

A) identical with at least a portion of the intrinsic endogenous B-hydroxylase transcript of plant and/or

B) at least a portion with the intrinsic endogenous B-hydroxylase promoter sequence of plant is identical.

In the method for the invention, in order to reduce endogenous B-hydroxylase activity, preferably RNA is imported plant, wherein said RNA contains tool duplex structure zone and contain following nucleotide sequence in this zone

Think that term " endogenous B-hydroxylase transcript " consciousness is meant the part of transcribing of endogenous Beta-hydroxylase gene, wherein except the sequence of coding endogenous B-hydroxylase, also comprises for example non-coding sequence such as UTR.

Think that the RNA of " identical with at least a portion of the intrinsic endogenous B-hydroxylase promoter sequence of plant " is meant that preferably the theoretical transcript of RNA sequence and endogenous B-hydroxylase promoter sequence refers to that just its corresponding RNA sequence is to the identical fact of small part.

Think that " part " meaning of the endogenous B-hydroxylase promoter sequence that endogenous B-hydroxylase transcript that plant is intrinsic or plant are intrinsic is meant the partial sequence from several base-pairs to the as many as full sequence in transcript or the promoter sequence.By those skilled in the art by the routine test optimal length of deciding section sequence easily.

Preferably, find out partial sequence to obtain the mode that the high as far as possible specificity and the activity of other enzymes (not expecting its active reduction) do not reduce.Therefore for the partial sequence of endogenous B-hydroxylase-dsRNA, the partial sequence that is chosen in non-existent part endogenous B-hydroxylase transcript in other activity and/or endogenous B-hydroxylase promoter sequence is favourable.

Therefore, in particularly preferred embodiments, the part of the intrinsic endogenous B-hydroxylase transcript of the sequence that contains of endogenous B-hydroxylase-dsRNA and plant is identical and contain 5 ' terminal or 3 ' end of the nucleic acid of the intrinsic coding endogenous B-hydroxylase of plant.Especially, transcript 5 ' or 3 ' non-translational region are suitable for producing the selectivity duplex structure.

The invention further relates to double stranded rna molecule (dsRNA molecule), when it is imported plant (or its cell, tissue, organ or propagating materials), cause that the endogenous B-hydroxylase reduces.

(the endogenous B-hydroxylase-dsRNA), this double stranded rna molecule preferably comprises to the invention further relates to the double stranded rna molecule that reduces the expression of endogenous B-hydroxylase

A) contain at least a identical with at least a portion of " having a mind to " RNA-endogenous B-hydroxylase transcript basically ribonucleotide acid sequence " having a mind to " RNA chain and

In order to transform plant, preferably used the nucleic acid construct that it is imported plant and be transcribed into endogenous B-hydroxylase-dsRNA in plant with endogenous B-hydroxylase-dsRNA.

The invention still further relates in addition and can be transcribed into following nucleic acid construct:

B) basically preferably fully and " antisense " RNA chain of having a mind to the chain complementation of the RNA a).

These nucleic acid constructs are also referred to as expression cassette or expression vector hereinafter.

About the dsRNA molecule, think that endogenous B-hydroxylase nucleotide sequence or its corresponding transcript preferably are meant the sequence according to SEQ ID NO:139 or its part.

" substantially the same " thus the meaning be meant that comparing the dsRNA sequence with endogenous B-hydroxylase target sequence also can have effective reduction that insertion, disappearance and single point mutation cause expression.Preferably, " having a mind to " chain of inhibition dsRNA and endogenous Beta-hydroxylase gene to small part " is had a mind to " between the rna transcription basis or the homology between " antisense " chain and the endogenous Beta-hydroxylase gene complementary strand is at least 75%, be preferably at least 80%, very particularly preferably be at least 90%, be most preferably 100%.

Reduce for causing that the endogenous B-hydroxylase is expressed effectively, it is not absolute necessary having 100% sequence homogeneity between dsRNA and the endogenous Beta-hydroxylase gene transcript.Therefore, tolerance is because genetic mutation, polymorphism or evolutionary divergence and method that the sequence that may exist departs from is favourable.It is possible for example using dsRNA from a kind of organism endogenous B-hydroxylase sequence to suppress in the another kind of organism that the endogenous B-hydroxylase expresses.For this purpose, dsRNA preferably contains the sequence area corresponding to the endogenous Beta-hydroxylase gene transcript of conserved region.Can obtain described conserved region fast by sequence alignment.

In addition, the dsRNA of " substantially the same " also can be defined as can with the nucleotide sequence of a part of endogenous Beta-hydroxylase gene transcript hybridization (for example in 400mM NaCl, 40mM PIPES pH 6.4,1mM EDTA 50 ℃ or 70 ℃ of hybridization 12-16 hour).

In another embodiment, endogenous B-hydroxylase-dsRNA comprises

A) contain at least a basically with endogenous Beta-hydroxylase gene promoter region " have a mind to " rna transcription this to " having a mind to " RNA chain of the identical ribonucleotide acid sequence of small part and

The corresponding nucleic construct that is preferably used for transforming plant comprises

A) identical with at least a portion of endogenous Beta-hydroxylase gene promoter region basically " having a mind to " DNA chain and

B) basically preferably fully and " antisense " DNA chain of " having a mind to " the chain complementation of the DNA a).

In order to produce the endogenous B-hydroxylase sequence that is used to reduce endogenous B-hydroxylase activity,, particularly preferably use following partial sequence particularly for marigold:

SEQ ID NO:141: the fragment intentionally of endogenous B-hydroxylase 5 ' stub area

SEQ ID NO:142: the antisense fragment of endogenous B-hydroxylase 5 ' stub area

DsRNA can be made up of one or more polybribonucleotide chain.Certainly, in order to realize identical purpose, also can be with multiple different dsRNA molecule transfered cell or organism, each in them all contains one of ribonucleotide acid sequence part defined above.

By the independent RNA chain of two complementations or preferably can form double-stranded dsRNA structure by the RNA start of chain of single self complementation.In this case, " have a mind to " RNA chain and " antisense " RNA chain preferably with the form of oppositely " repetition " covalent bond each other.

For example, dsRNA also can contain hairpin structure described in WO 99/53050, by with catenation sequence (" joint "; Introne for example) connects " having a mind to " and " antisense " chain.The dsRNA structure of self complementation is preferred, and this is because they only need the complementary RNA chain of molar ratios such as expressing a RNA sequence and always contain.Preferably, catenation sequence is that introne is (for example from the introne of potato ST-LS1 gene; (1990) Mol Gen Genet220 (2) such as Vancanneyt GF: 245-250).

Other preferred embodiment that reduces endogenous B-hydroxylase activity is similar to the above-mentioned preferred embodiment that is used to reduce ε-cyclase activity, has just wherein replaced ε-cyclase with the endogenous B-hydroxylase.

In the methods of the invention, particularly preferably use has the genetically modified plant that has made up following genetic modification:

Compare with wild type, it is active and have a genetically modified plant of the hydroxylase activity that has improved to have the ketolase that improved or induce in floral leaf,

Compare with wild type, in floral leaf, have the genetically modified plant that the ketolase that improved or induce is active and have the beta cyclase activity that has improved,

Compare with wild type, it is active and have a genetically modified plant of the ε-cyclase activity that has reduced to have the ketolase that improved or induce in floral leaf,

Compare with wild type, it is active and have a hydroxylase activity that has improved and the genetically modified plant of the beta cyclase activity that improved to have the ketolase that improved or induce in floral leaf,

Compare with wild type, it is active and have a hydroxylase activity that has improved and the genetically modified plant of ε-cyclase activity of having reduced to have the ketolase that improved or induce in floral leaf,

Compare with wild type, it is active and have a genetically modified plant of the active and ε-cyclase activity that reduced of the beta cyclase that improved to have the ketolase that improved or induce in floral leaf, and

Compare with wild type, in floral leaf, have the ketolase that improved or induce active and have the hydroxylase activity that improved, the beta cyclase that improved is active and the genetically modified plant of ε-cyclase activity of having reduced,

Compare with wild type, the genetically modified plant of ε-cyclase activity of in floral leaf, have the ketolase activity that improved or induce, having reduced, the beta cyclase activity that improved,

Compare with wild type, the genetically modified plant of the ε-cyclase activity that in floral leaf, have the ketolase activity that improved or induce, reduced and the endogenous B-hydroxylase activity that reduced,

Compare with wild type, the genetically modified plant of the ε-cyclase activity that in floral leaf, have the ketolase activity that improved or induce, reduced and the hydroxylase activity that improved,

Compare with wild type, the genetically modified plant of the active and hydroxylase activity that improved of the beta cyclase that in floral leaf, have the ketolase activity that improved or induce, improved,

Compare with wild type, the genetically modified plant of the active and endogenous B-hydroxylase activity that reduced of the beta cyclase that in floral leaf, have the ketolase activity that improved or induce, improved,

Compare with wild type, in floral leaf, have the genetically modified plant of the active and beta cyclase activity that improved of the ketolase that improved or induce,

Compare with wild type, in floral leaf, has the ketolase activity that has improved or induce, ε-the cyclase activity that reduced and at least a activity that other has improved are selected from the HMG-CoA reductase activity, (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity, 1-deoxidation-D-wood sugar-5-phosphate synthase activity, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity, isopentene group-bisphosphate Δ-isomerase activity, Mang ox base-bisphosphate synthase activity, method acyl-bisphosphate synthase activity, Mang ox base Mang ox base bisphosphate synthase activity, the phytoene synthase activity, the phytoene desaturase activity, sigma carotene desaturase activity, the crtlSO activity, the genetically modified plant of FtsZ activity or MinD activity

Compare with wild type, ε-cyclase activity of have the ketolase activity that improved or induce in floral leaf, having reduced, the beta cyclase that has improved is active and the genetically modified plant of the hydroxylase activity that improved,

Compare with wild type, ε-cyclase activity of have the ketolase activity that improved or induce in floral leaf, having reduced, the beta cyclase that has improved is active and the genetically modified plant of the endogenous B-hydroxylase activity that reduced,

Compare with wild type, the genetically modified plant of the ε-cyclase activity that in floral leaf, have the ketolase activity that improved or induce, reduced and the beta cyclase activity that improved,

Compare with wild type, the genetically modified plant of ε-cyclase activity of in floral leaf, have the ketolase activity that improved or induce, having reduced, the hydroxylase activity that has improved and the endogenous B-hydroxylase activity that reduced,

Compare with wild type, the genetically modified plant of the beta cyclase activity that in floral leaf, have the ketolase activity that improved or induce, improved, the hydroxylase activity that has improved and the endogenous B-hydroxylase activity that reduced,

Compare with wild type, in floral leaf, has the ketolase activity that has improved or induce, ε-the cyclase activity that has reduced, active and at least a activity that other has improved of the beta cyclase that has improved is selected from the HMG-CoA reductase activity, (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity, 1-deoxidation-D-wood sugar-5-phosphate synthase activity, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity, isopentene group-bisphosphate D-isomerase activity, Mang ox base-bisphosphate synthase activity, method acyl-bisphosphate synthase activity, Mang ox base Mang ox base bisphosphate synthase activity, the phytoene synthase activity, the phytoene desaturase activity, sigma carotene desaturase activity, the crtlSO activity, the genetically modified plant of FtsZ activity or MinD activity

Compare with wild type, the genetically modified plant of ε-cyclase activity of in floral leaf, have the ketolase activity that improved or induce, having reduced, the beta cyclase activity that has improved, the hydroxylase activity that has improved and the endogenous B-hydroxylase activity that reduced

Compare with wild type, in floral leaf, has the ketolase activity that has improved or induce, ε-the cyclase activity that has reduced, the hydroxylase activity that improved and at least a activity that other has improved are selected from the HMG-CoA reductase activity, (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity, 1-deoxidation-D-wood sugar-5-phosphate synthase activity, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity, isopentene group-bisphosphate D-isomerase activity, hold together Niu Erji-bisphosphate synthase activity, method acyl-bisphosphate synthase activity, Mang ox base Mang ox base bisphosphate synthase activity, the phytoene synthase activity, the phytoene desaturase activity, sigma carotene desaturase activity, the crtlSO activity, the genetically modified plant of FtsZ activity or MinD activity

Compare with wild type, in floral leaf, has the ketolase activity that has improved or induce, ε-the cyclase activity that has reduced, active and at least a activity that other has improved of the endogenous B-hydroxylase that has reduced is selected from the HMG-CoA reductase activity, (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity, 1-deoxidation-D-wood sugar-5-phosphate synthase activity, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity, isopentene group-bisphosphate D-isomerase activity, Mang ox base-bisphosphate synthase activity, method acyl-bisphosphate synthase activity, Mang ox base Mang ox base bisphosphate synthase activity, the phytoene synthase activity, the phytoene desaturase activity, sigma carotene desaturase activity, the crtlSO activity, the genetically modified plant of FtsZ activity or MinD activity

Compare with wild type, in floral leaf, has the ketolase activity that has improved or induce, the beta cyclase activity that has improved, the hydroxylase activity that improved and at least a activity that other has improved are selected from the HMG-CoA reductase activity, (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity, 1-deoxidation-D-wood sugar-5-phosphate synthase activity, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity, isopentene group-bisphosphate Δ-isomerase activity, Mang ox base-bisphosphate synthase activity, method acyl-bisphosphate synthase activity, Mang ox base Mang ox base bisphosphate synthase activity, the phytoene synthase activity, the phytoene desaturase activity, sigma carotene desaturase activity, the crtlSO activity, the genetically modified plant of FtsZ activity or MinD activity

Compare with wild type, in floral leaf, has the ketolase activity that has improved or induce, the beta cyclase activity that has improved, active and at least a activity that other has improved of the endogenous B-hydroxylase that has reduced is selected from the HMG-CoA reductase activity, (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity, 1-deoxidation-D-wood sugar-5-phosphate synthase activity, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity, isopentene group-bisphosphate D-isomerase activity, Mang ox base-bisphosphate synthase activity, method acyl-bisphosphate synthase activity, Mang ox base Mang ox base bisphosphate synthase activity, the phytoene synthase activity, the phytoene desaturase activity, sigma carotene desaturase activity, the crtlSO activity, the genetically modified plant of FtsZ activity or MinD activity

Compare with wild type, ε-cyclase activity of have the ketolase activity that improved or induce in floral leaf, having reduced, the beta cyclase that has improved is active and the genetically modified plant of the hydroxylase activity that improved and the B-hydroxylase activity that reduced,

Compare with wild type, in floral leaf, has the ketolase activity that has improved or induce, ε-the cyclase activity that has reduced, the beta cyclase activity that has improved, the hydroxylase activity that improved and at least a activity that other has improved are selected from the HMG-CoA reductase activity, (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity, 1-deoxidation-D-wood sugar-5-phosphate synthase activity, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity, isopentene group-bisphosphate D-isomerase activity, Mang ox base-bisphosphate synthase activity, method acyl-bisphosphate synthase activity, Mang ox base Mang ox base bisphosphate synthase activity, the phytoene synthase activity, the phytoene desaturase activity, sigma carotene desaturase activity, the crtlSO activity, the genetically modified plant of FtsZ activity or MinD activity

Compare with wild type, in floral leaf, have the ketolase activity that has improved or induce, ε-the cyclase activity that has reduced, the beta cyclase activity that has improved, active and at least a activity that other has improved of the endogenous B-hydroxylase that has reduced is selected from the HMG-CoA reductase activity, (E)-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity, 1-deoxidation-D-wood sugar-5-phosphate synthase activity, 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity, isopentene group-bisphosphate D-isomerase activity, Mang ox base-bisphosphate synthase activity, method acyl-bisphosphate synthase activity, Mang ox base Mang ox base bisphosphate synthase activity, the phytoene synthase activity, the phytoene desaturase activity, sigma carotene desaturase activity, the crtlSO activity, the genetically modified plant of FtsZ activity or MinD activity.

Especially preferably compare with wild type, the genetically modified plant of the active and hydroxylase activity that improved of the beta cyclase that in floral leaf, have the ketolase activity that improved or induce, improved,

Induce the ketolase activity of raising by importing such nucleic acid, the protein of described nucleic acid coding comprises amino acid sequence SEQ ID NO:2 or by this sequence is carried out the sequence that amino acid replacement, insertion or disappearance obtain, and the aftermentioned sequence has at least 20% homogeneity with sequence SEQ ID NO:2 and has the enzymatic property of ketolase on amino acid levels.

Induce the beta cyclase activity of raising by importing such nucleic acid, the beta cyclase of described nucleic acid coding comprises amino acid sequence SEQ ID NO:110 or by this sequence is carried out the sequence that amino acid replacement, insertion or disappearance obtain, the aftermentioned sequence has at least 20% homogeneity with sequence SEQ ID NO:110 on amino acid levels.

Induce the hydroxylase activity of raising by importing such nucleic acid, the hydroxylase of described nucleic acid coding comprises amino acid sequence SEQ ID NO:108 or by this sequence is carried out the sequence that amino acid replacement, insertion or disappearance obtain, the aftermentioned sequence has at least 20% homogeneity with sequence SEQ ID NO:108 on amino acid levels.

Especially preferably compare with wild type, the genetically modified plant of ε-cyclase activity of in floral leaf, have the ketolase activity that improved or induce, having reduced, the beta cyclase activity that has improved, the hydroxylase activity that has improved and the endogenous B-hydroxylase activity that reduced

Induce the hydroxylase activity of raising by importing such nucleic acid, the hydroxylase of described nucleic acid coding comprises amino acid sequence SEQ ID NO:108 or by this sequence is carried out the sequence that amino acid replacement, insertion or disappearance obtain, the aftermentioned sequence has at least 20% homogeneity with sequence SEQ ID NO:108 on amino acid levels.And the endogenous B-hydroxylase activity of inducing the ε-cyclase activity that has reduced and having reduced according to above-mentioned embodiment preferred.

Described in hereinafter, for example, can produce these genetically modified plants of Tagetes by importing independent nucleic acid construct (expression cassette) or containing the multiple construct of two, three or four described activity of as many as by importing.

Hereinafter, mode is described in the generation of the genetically modified plant that has the ketolase activity that has improved or induce in the floral leaf by way of example.β-HMG-CoA/ ( E )-4--3--2--/1--D--5-/1--D--5-/-D-/-/-////ζ-/crtlSO/FtsZ/MinD； Can realize the raising of other activity, for example hydroxylase activity and/or beta cyclase activity and/or HMG-CoA reductase activity and/or ( E )-4-hydroxy-3-methyl but-2-ene base-bisphosphate reductase activity and/or 1-deoxidation-D-wood sugar-5-phosphate synthase activity and/or 1-deoxidation-D-wood sugar-5-phosphoric acid reduction isomerase activity and/or isopentene group-bisphosphate D-isomerase activity and/or Mang ox base-bisphosphate synthase activity and/or method acyl-bisphosphate synthase activity and/or Mang ox base Mang ox base bisphosphate synthase activity and/or phytoene synthase activity and/or phytoene desaturase activity and/or sigma carotene desaturase activity and/or crtlSO activity and/or FtsZ activity and/or MinD are active. Use the reverse repetitive nucleic acid sequence of ε-cyclase anti sense nucleotide sequence or the reverse repetitive nucleic acid sequence of ε-cyclase or use endogenous B-hydroxylase anti sense nucleotide sequence or endogenous B-hydroxylase to replace the nucleotide sequence of coding ketolase in a similar manner; Can realize for example reduction of ε-cyclase activity or endogenous B-hydroxylase activity of other enzymatic activity.Making up under the situation of genetic modification, implementing discriminably to transform or implement to transform by multiple construct.

Preferably, contain above-mentioned coding ketolase and the functional nucleic acid construct that is connected in the nucleic acid on one or more adjustment signals of guaranteeing in plant, to transcribe and translate by use and carry out the conversion of initial plant, produce the Tagetes genetically modified plants.

Wherein functional these nucleic acid constructs that are connected on one or more adjustment signals of guaranteeing to transcribe in plant and translate of nucleic acid sequence encoding are also referred to as expression cassette hereinafter.

Preferably, adjustment signal contains one or more promoters of transcribing and translating guaranteed in plant.

Expression cassette contains adjustment signal, that is to say the modulability nucleotide sequence that coded sequence is expressed in the control host cell.According to embodiment preferred, expression cassette comprises upstream (that is to say at coded sequence 5 ' end), promoter, downstream (that is to say at 3 ' end), polyadenylation signal and also comprises other as required and can be operatively connected in the controlling element of the coded sequence of at least one said gene.Can be operatively connected the meaning be meant promoter, coded sequence, terminator and as required other controlling element mode that can both in coded sequence is expressed, fulfil its function in a suitable manner with each controlling element arrange in proper order.

Hereinafter, describe preferred nucleic acid construct, expression cassette and carrier that is used for plants of tagetes species and the method that is used to produce the Tagetes genetically modified plants by way of example, also had Tagetes genetically modified plants self.

The sequence that is preferred for being operatively connected is that (but being not limited to) guarantees the target sequence of Subcellular Localization in apoplast, vacuole, plastid, mitochondria, endoplasmic reticulum (ER), nucleus, oil body or other compartment and such as the translational enhancer (Gallie etc. from tobacco mosaic virus (TMV) (tobacco mosaic virus) 5 ' targeting sequencing, Nucl.Acids Res.15 (1987), 8693-8711).

In principle, can control any promoter that foreign gene expresses in plant and all be suitable for promoter as expression cassette.

" composing type " promoter be meant those majority preferably the very long in a organized way development of plants phase preferably guarantee expression promoter at the All Time point of development of plants.

Especially preferably use plant promoter or come from the promoter of plant virus.Particularly, the promoter of CaMV cauliflower mosaic virus (CaMV cauliflower mosaic virus) 35S transcript is preferred (Franck etc. (1980) Cell 21:285-294; Odell etc. (1985) Nature313:810-812; Shewmaker etc. (1985) Virology 140:281-288; Gardner etc. (1986) Plant Mol Biol 6:221-228) or 19S CaMV promoter (US 5,352,605; WO 84/02913; Benfey etc. (1989) EMBO J 8:2195-2202).

Other suitable constitutive promoter is that (US 4 for promoter for pds promoter (Pecker etc. (1992) Proc.Natl.Acad.Sci USA 89:4962-4966) or " rubisco small subunit (SSU) ", 962,028), legumin B promoter (GenBank accession number X03677), nopaline synthase promoter from Agrobacterium, the TR double-promoter, OCS (octopine synthase) promoter from Agrobacterium, ubiquitin promoter (HoltorfS etc. (1995) PlantMol Biol 29:637-649), ubiquitin 1 promoter (Christensen etc. (1992) Plant Mol Biol18:675-689; Bruce etc. (1989) Proc Natl Acad Sci USA 86:9692-9696), the Smas promoter, (US 5 for the cinnamyl-alcohol dehydrogenase promoter, 683,439), vacuole ATP enzyme subunit promoter or from promoter (WO 91/13991), the Pnit promoter (Y07648.L of the protein that is rich in proline of wheat, Hillebrand etc. (1998), Plant.Mol.Biol.36,89-99, Hillebrand etc. (1996), Gene, 170,197-200) and other well known by persons skilled in the art in plant the promoter of the gene of constitutive expression.

But expression cassette also can comprise the promoter (survey article: Gatz etc. (1997) Annu Rev Plant Physiol Plant Mol Biol 48:89-108) of chemistry induction type, by chemical inducible promoter can be in special time point control plant the ketolase expression of gene.Can use this type of promoter equally, for example PRP1 promoter (Ward etc. (1993) Plant Mol Biol 22:361-366), salicylic acid-inducible promoter (WO 95/19443), benzsulfamide-inducible promoter (EP 0 388 186), tetracycline-inducible promoter (Gatz etc. (1992) Plant J 2:397-404), abscisic acid-inducible promoter (EP 0 335 528) and ethanol-or cyclohexanone-inducible promoter (WO 93/21334).

And, the promoter of being induced by biology or abiotic stress also is preferred, for example pathogen-the inducible promoter of PRP1 gene (Ward etc. (1993) Plant Mol Biol 22:361-366), (US 5 from heat-derivable hsp70 of tomato or hsp80 promoter, 187,267), from cold-derivable AMS promoter (WO 96/12814), light-derivable PPDK promoter or wound-derivable pinII promoter (EP 375091) of potato.

Pathogen-inducible promoter comprises the result who attacks as pathogen and the promoter of the gene of inducing, described gene is PR albumen, SAR albumen, β-1 for example, ((1983) Neth J Plant Pathol 89:245-254 such as Redolfi for example such as the gene of 3-dextranase, chitinase etc.; Uknes etc. (1992) The Plant Cell 4:645-656; Van Loon (1985) Plant Mol Viral4:111-116; Marineau etc. (1987) Plant Mol Biol 9:335-342; Matton etc. (1987) Molecular Plant-Microbe Interactions 2:325-342; Somssich etc. (1986) ProcNatl Acad Sci USA 83:2427-2430; Somssich etc. (1988) Mol Gen Genetics2:93-98; Chen etc. (1996) Plant J 10:955-966; Zhang and Sing (1994) ProcNatl Acad Sci USA 91:2507-2511; Warner etc. (1993) Plant J 3:191-201; Siebertz etc. (1989) Plant Cell 1:961-968 (1989).

Wound-the inducible promoter that also comprises some genes, described gene such as pinII gene (Ryan (1990) Ann Rev Phytopath 28:425-449; Duan etc. (1996) Nat Biotech14:494-498), (US 5 for wun1 and wun2 gene, 428,148), win1 and win2 gene (Stanford etc. (1989) Mol Gen Genet 215:200-208), systemin (McGurl etc. (1992) Science225:1570-1573), WIP1 gene (Rohmeier etc. (1993) Plant Mol Biol 22:783-792; Ekelkamp etc. (1993) FEBS Letters 323:73-76), MPI gene (Corderok etc. (1994) The Plant J 6 (2): 141-150) or the like.

Other suitable promoter is for example fruit maturation specific promoter, as the fruit maturation specific promoter (WO 94/21794, and EP 409 625) from tomato.Certainly, because different tissues forms to grow the dependence mode, grow the dependence promoter and also comprise the portion of tissue specific promoter.

In addition, especially preferably guarantee in tissue or plant part, to carry out those promoters of for example keto-acid carotenoid biosynthesis or its precursor generation expression.For example preferably have the promoter of flower pesticide, ovary, petal, sepal, flower, leaf, stem and root-specific and their combination.

Stem tuber-, storage root-or root-specific promoter be for example, to start the promoter of subclass I (B33) or cathepsin D's inhibitor from the patatin of potato.

The leaf specific promoter be for example from the promoter (WO 97/05900) of the cytosol FBP enzyme of potato, from the SSU promoter (small subunit) or the ST-LSI promoter (Stockhaus etc. (1989) EMBO J 8:2445-2451) of the rubisco (ribulose-1,5-bisphosphate, 5-diphosphonic acid carboxylase) of potato.

The flower specific promoter is for for example from phytoene synthase promoter (WO 92/16635) or P-rr gene promoter (WO 98/22593) or the AP3 promoter (seeing embodiment 1) of mouseearcress.

The flower pesticide specific promoter is for example 5126 promoters (US 5,689,049, and US 5,689,051), glob-l promoter or g-zeins promoter.

Other are suitable for, and expression promoter is described in (1987) Meth inEnzymol 153:253-277 such as Rogers in plant; (1989) Proc Natl Acad Sci USA:8402-8406 such as Schardl etc. (1987) Gene 61:1-11 and Berger).

Promoter described in all the application makes that usually expressing ketolase in the floral leaf of plant of the present invention becomes possibility.

Particularly preferably being composing type in the inventive method spends special and the special promoter of floral leaf especially.

Reorganization and clone technology according to routine, for example be described in T.Maniatis, E.F.Fritsch and J.Sambrook, Molecular Cloning:A Laboratory Manual, Cold SpringHarbor Laboratory, Cold Spring Harbor, NY (1989), T.J.Silhavy, M.L.Berman and L.W.Enquist, Experiments with Gene Fusions, ColdSpring Harbor Laboratory, Cold Spring Harbor, NY (1984) and Ausubel, F.M. etc., Current Protocols in Molecular Biology, reorganization and clone technology among the Greene Publishing Assoc.and Wiley-Interscience (1987) preferably produce expression cassette by nucleic acid that merges suitable promoter and above-mentioned coding ketolase and nucleic acid and the polyadenylation signal that preferably inserts coding plastid unitransport peptide between promoter and this nucleotide sequence.

Preferably insert and the nucleic acid of the plastid transit peptides of encoding guarantees to be positioned plastid, and particularly be positioned chromoplast.

Also can use the expression cassette of its nucleic acid sequence encoding ketolase fused protein, a part of fused protein is as the transit peptides of control polypeptide transposition.For the special transit peptides of chromoplast is preferred, and special transit peptides gets off from ketolase is partially digested after the ketolase transposition enters chromoplast.

Particularly preferred transit peptides is from the transit peptides of tobacco plastid transketolase or other transit peptides (for example transit peptides of rubisco (rbcS) small subunit or ferredoxin NADP oxidoreducing enzyme and isopentenylpyrophosphate isomerase-2 or its function equivalent).

Nucleotide sequence from three boxes of the plastid transit peptides of tobacco plastid transketolase is particularly preferred, and they have three reading frames of ATG codon at the NcoI restriction enzyme site as the KpnI/BamHI fragment:

pTP09

KpnI_GGTACCATGGCGTCTTCTTCTTCTCTCACTCTCTCTCAAGC

TATCCTCTCTCGTTCTGTCCCTCGCCATGGCTCTGCCTCTTCTTC

TCAACTTTCCCCTTCTTCTCTCACTTTTTCCGGCCTTAAATCCAA

TCCCAATATCACCACCTCCCGCCGCCGTACTCCTTCCTCCGCCG

CCGCCGCCGCCGTCGTAAGGTCACCGGCGATTCGTGCCTCAGCT

GCAACCGAAACCATAGAGAAAACTGAGACTGCGGGATCC_BamH

I

pTP10

KpnI_GGTACCATGGCGTCTTCTTCTTCTCTCACTCTCTCTCAAGC

TATCCTCTCTCGTTCTGTCCCTCGCCATGGCTCTGCCTCTTCTTC

TCAACTTTCCCCTTCTTCTCTCACTTTTTCCGGCCTTAAATCCAA

TCCCAATATCACCACCTCCCGCCGCCGTACTCCTTCCTCCGCCG

CCGCCGCCGCCGTCGTAAGGTCACCGGCGATTCGTGCCTCAGCT

GCAACCGAAACCATAGAGAAAACTGAGACTGCGCTGGATCC_Ba

mHI

pTP11

KpnI_GGTACCATGGCGTCTTCTTCTTCTCTCACTCTCTCTCAAGC

TATCCTCTCTCGTTCTGTCCCTCGCCATGGCTCTGCCTCTTCTTC

TCAACTTTCCCCTTCTTCTCTCACTTTTTCCGGCCTTAAATCCAA

TCCCAATATCACCACCTCCCGCCGCCGTACTCCTTCCTCCGCCG

CCGCCGCCGCCGTCGTAAGGTCACCGGCGATTCGTGCCTCAGCT

GCAACCGAAACCATAGAGAAAACTGAGACTGCGGGGATCC_Bam

HI

Other example of plastid transit peptides is from the transit peptides of the isopentenylpyrophosphate isomerase-2 (IPP-2) of mouseearcress plastid with from the transit peptides (Guerineau of pea carboxydismutase small subunit (rbcS), F, Woolston, S, Brooks, L, Muuineaux, P (1988) is used for the expression cassette with foreign protein target chromoplast, Nucl.Acids Res.16:11380).

Nucleic acid of the present invention can synthesize preparation or natural generation or can contain synthetic and natural acid mixture of ingredients, and also can partly be made up of multiple organic multiple heterologous gene.

As mentioned above, have by plants of tagetes species the synthesizing ribonucleotide sequence of preferred codon be preferred.Can determine preferred these codons of plant from the codon that most purpose plant species, has high expressed protein frequency.

In the preparation of expression cassette, can operate multiple dna fragmentation and obtain with the correct direction easy-to-read and place the nucleotide sequence of correct reading frame.For being connected to each other of dna fragmentation, joint or connexon can be added on the fragment.

Easily, can the joint of one or more restriction enzyme sites or polylinker be will contain with transcriptional orientation and promoter and terminator zone are provided in to insert described sequence.Usually, joint has from 1 to 10, and usually from 1 to 8, from 2 to 6 restriction enzyme sites preferably.Usually, in the regulation and control zone, the joint size is less than 100bp, is less than 60bp usually, but is at least 5bp.For host plant, promoter can be natural or homology also can be external or allos.Expression cassette 5 '-3 ' transcriptional orientation preferably contains promoter, nucleic acid sequence encoding or nucleic acid construct and tanscription termination zone.Multiple termination zone can be replaced as required each other.

The example of terminator is 35S terminator (Guerineau etc. (1988) Nucl Acids Res.16:11380), no terminator (Depicker A, Stachel S, Dhaese P, Zambryski P, Goodman HM., nopaline synthase: transcripting spectrum and dna sequence dna, J Mol Appl Genet.1982; 1 (6): 561-73) or ocs terminator (Gielen, J, de Beuckeleer, M, Seurinck, J, Debroek, H, de Greve, H, Lemmers, M, van Montagu, M, Schell, J (1984), the TL-DNA total order of Agrobacterium tumefaciems plasmid pTiAch5, EMBO is J.3:835-846).

In addition, can use the operation that the restriction enzyme site that is complementary is provided or removes DNA redundant or restriction enzyme site.Insert, disappearance or alternative can use mutagenesis in vitro, " primer reparation ", restriction enzyme digestion as conversion during with transversion or be connected.

Use suitable operation, for example restriction enzyme digestion, " digestion " or mend flat jag and form blunt ends can obtain the complementary terminal of the fragment that is used to connect.

Preferred polyadenylation signal is the plant polyadenylation signal, preferably correspond essentially to T-DNA polyadenylation signal from Agrobacterium tumefaciems (Agrobacterium tumefaciens), especially Ti-plasmids pTiACH5 (Gielen etc., EMBO is (1984) J.3,835ff) polyadenylation signal or its function equivalent of T-DNA gene 3 (octopine synthase).

Foreign gene is transferred to Plant Genome is called conversion.

Therefore, can utilization itself known method transforms and from the plant tissue or the plant cell aftergrowth of instantaneous or stable conversion.

The proper method that is used for transforming plant is to take in the biology that carries out protoplast transformation, uses particle gun to launch (biolistic) method of hitting (being also referred to as the particle bombardment method), electroporation, hatch dry embryo method, microinjection and above-mentioned by agriculture bacillus mediated transgenosis containing dna solution by the polyethylene glycol inducing DNA.Described method is described in for example Transgenic Plants, 1 volume, B.Jenes among the Engineeringand Utilization (S.D.Kung and R.Wu Academic Press (1993) publish) etc., gene transfer technique, 128-143, and be described in Potrykus, Annu.Rev.Plant Physiol.Plant Molec.Biol.42 (1991), 205-225).

Preferably, the construct that desire is expressed is cloned into and is suitable for transforming in the carrier of Agrobacterium tumefaciems, pBin19 (Bevan etc., Nucl.Acids Res.12 (1984), 8711) or particularly preferably be pSUN2, pSUN3, pSUN4 or pSUN5 (WO 02/00900) for example.

The Agrobacterium that transforms by expression plasmid can be used to transform plant in known manner, for example by dipping bath wound leaf or vanes block in transforming Agrobacterium solution and cultivate them then in suitable culture medium.

Be also referred to as genetically modified plants hereinafter for preferably producing genetically modified plant, the amalgamation and expression box of expressing ketolase be cloned into carrier for example pBin19 or the particularly pSUN2 that is suitable for being transformed into Agrobacterium tumefaciems.To be used to transform plant with the Agrobacterium that this kind carrier transforms in known manner then, particularly by dipping bath wound leaf or vanes block in transforming Agrobacterium solution and in suitable culture medium, cultivate them then and obtain cultivating plant.

Carry out Plant Transformation by Agrobacterium and especially be disclosed in Transgenic Plants, Vol.1, Engineering and Utilization (S.D.Kung and R.Wu, Academic Press, 1993 publish) F.F.White in the book, be used for the carrier of transgenosis in the higher plant; The 15-38 page or leaf.In known manner, can bear again from the transformant of the leaf of wound or vanes block and contain the genetically modified plants of gene that are integrated into expression cassette and express the nucleic acid of coding ketolase.

For the conversion that transforms the Tagetes host plant with nucleic acid with coding ketolase, its expression cassette is integrated the insertion recombinant vector, described carrier DNA comprises extra functional adjustment signal, for example is used to the sequence of duplicating or integrating.Appropriate carriers especially is described in " Methods in PlantMolecular Biology and Biotechnology " (CRC Press), 6/7 chapter, 71-119 page or leaf (1993).

Use above-cited reorganization and clone technology, expression cassette can be cloned into appropriate carriers, for example amplification in Escherichia coli (E.coli.) to allow their amplification.Suitable cloning vector is pJIT117 (Guerineau etc. (1988) Nucl.Acids Res.16:11380), pBR332, pUC series, M13mp series and pACYC184 especially.Special appropriate carriers is the binary vector that not only can also can duplicate in Agrobacterium in Escherichia coli.

According to selection to promoter, can be in floral leaf composing type ground or be preferably specifically express.

Compare with wild type, the genetically modified plant of Tagetes of the present invention contains a certain amount of astaxanthin, especially in petal.

As mentioned above, the present invention relates to the purposes of the plants of tagetes species of astaxanthin-containing or its part or relate to the plants of tagetes species of astaxanthin-containing or the purposes of the astaxanthin-containing extract of its part, be used to be administered orally in animal.

In preferred embodiments, the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or its part or astaxanthin-containing or its part is used to make animal painted with corresponding animal product.

Think that the astaxanthin-containing extract of astaxanthin-containing plant or plant part preferably is meant the solution that contains the astaxanthin-containing that produces from astaxanthin-containing plant or plant part extracting by at least a appropriate solvent.According to employed solvent and employed chemistry and physical purification method, astaxanthin can be present in the extract with the purity of any expection.Suitably prepare the astaxanthin-containing plant before the extracting or plant part is favourable, for example dried plant or plant part and it is pulverized, it is chosen wantonly in proper order.

By organic solvent for example acetone, hexane, carrene, methyl tertiary butyl ether(MTBE), or by solvent mixture such as ethanol/hexane or acetone/hexane, can be from the plant or the plant part extracting astaxanthin of astaxanthin-containing, the dry in advance and/or pulverizing of the plant of astaxanthin-containing or plant part as required.By the different mixed proportion of solvent, owing to have different polarity, extraction efficiency can be different.By this kind extracting, can be with high concentration enrichment astaxanthin.

Extract the purity that astaxanthin can further improve astaxanthin by vibration and chromatography mixture then.The common conduct list of astaxanthin-and the existence of two ester admixtures, exist with palmitate usually.

Compare according to it is considered herein that " painted " preferably is meant with the non-staining animal, at least a portion color of painted animal or animal product is strengthened or the generation color.The astaxanthin-containing pigment is painted and initial usually or strengthen pink to pink-redness.

Can painted preferred animal be the animal that is selected from fish, crustacean or birds by dosage forms for oral administration of the present invention, especially Galliformes (galliformes) and Anatridae.

Preferred fish are salmonid (salmonid), especially salmon or trout.

Preferred crustacean is shrimp and crab.

Preferred Galliformes is chicken, duck or goose.

Preferred Anatridae is flamingo (flamingo).

Depend on painted animal, preferably, think that painted animal product refers in particular to salmon or trout flesh of fish, the skin of chicken, duck or goose, the feather of chicken, duck, goose or flamingo, and ovum of chicken, duck or goose or yolk.

By directly or by dosage forms for oral administration in advance the plants of tagetes species of astaxanthin-containing or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part being mixed in animal feed goods wherein, can realize the plants of tagetes species of astaxanthin-containing or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part are administered orally in animal.

In preferred embodiments, the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part is mixed in the animal feed goods and with the animal feed goods be administered orally in animal.

Before being mixed in the animal feed goods, it is favourable that the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part is processed into a kind of form that can be mixed in corresponding animal feed goods and preferably produce high stability and high bioavilability astaxanthin in use field separately.

According to it being carried out the animal of dosage forms for oral administration and also according to the animal feed goods, therefore multiple procedure of processing can be favourable to its purpose.

For the plants of tagetes species or the plant part of astaxanthin-containing, in this embodiment, dry and/or pulverize the plant of astaxanthin-containing or plant part especially capitulum or petal are favourable.Particularly preferably, the plants of tagetes species of astaxanthin-containing or plant part exist with powder type.

Yet no matter astaxanthin-containing plants of tagetes species or plant part that each embodiment is prepared are processed or are not processed, and all can be mixed in the animal feed goods in a manner known way.

For the plants of tagetes species of astaxanthin-containing or the astaxanthin-containing extract of plant part, in this embodiment, multiple procedure of processing all is favourable.

If still the solvent of Cun Zaiing is on the physiology when harmless for corresponding animal, the astaxanthin-containing extract can directly be mixed in the animal feed goods.

Evaporate after the solvent that still exists, extract can use with the form of astaxanthin-containing powder or oil.

For example, resulting astaxanthin-containing powder or oil can mix fish oil, are applied to for example wheat flour or wipe broken marigold petal of dust carrier material, perhaps are included in alginates, gelatin or the lipid.

Therefore preferably liquid or powder type of the extract after astaxanthin-containing extract or the processing.

Yet no matter the astaxanthin-containing extract of the plants of tagetes species of the astaxanthin-containing that each embodiment is prepared or plant part is processed or is not processed, and all can be mixed in the animal feed goods in a manner known way.

Therefore the invention still further relates to the animal feed goods of the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species that comprises astaxanthin-containing or plant part or astaxanthin-containing or plant part.

The invention further relates to by plants of tagetes species or the astaxanthin-containing extract of plant part and the method for traditional animal feed combination results animal feed goods plants of tagetes species or the plant part or the astaxanthin-containing of astaxanthin-containing.

The method embodiment preferred be included in animal feed combination before, with the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part be processed into a kind of can with the form of animal feed combination.

For example, for fish, the fish feed goods can comprise other common fish feed composition, for example fish meal and/or other protein, oil (for example fish oil), cereal, vitamin, mineral matter, anticorrisive agent and the medicine that also contains convention amount as required.

For example, the general fish feed goods that are used for trout are grouped into by following one-tenth:

Composition	% weight	For 500kg body weight kg
Composition	% weight	For 500kg body weight kg	The wheaten starch of fish meal full-fat bean gel in advance	????30.00 ????20.00 ????18.00	?150.00 ?100.00 ?90.00

Premix vitamin Lipotril (50%) wheat gluten Sipernat 50S fish oil

????0.80 ????0.20 ????20.00 ????3.00 ????8.00

????4.00 ????1.00 ????100.00 ????15.00 ????40.00

For example, the general fish feed goods that are used for salmon are grouped into by following one-tenth:

Composition	% weight
Composition	% weight	Fish meal phytoprotein cereal vitamin/mineral antioxidant/anticorrisive agent fish oil	????75.00 ????5.00 ????7.80 ????1.00 ????0.20 ????11.00

In one embodiment, the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing extract preferably are mixed in the animal feed goods with the powder type of drying and pulverizing.

About fish feed, the animal feed goods of the plants of tagetes species of the resulting plants of tagetes species that contains astaxanthin-containing or plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part can be for example in a manner known way by granulating or particularly advantageously be extruded.

In preferred embodiments, the astaxanthin-containing extract mixes in the animal feed goods, preferably mixes with liquid form.This is favourable, particularly when producing the fish feed goods of extruding.Extrusion process for sensitive materials for example astaxanthin cause squeeze pressure, this may cause astaxanthin loss.Squeeze pressure mainly is the effect of mechanical force (kneading, shearing, compressing etc.), but also comprises by adding the hydro-thermal pressure that entry and steam cause, and also can observe oxidative pressure.

For fear of resultant astaxanthin loss, after extruding or dry run, under vacuum condition, can pass through PPA (use the back of granulating) technology using liquid astaxanthin-containing extract as above-mentioned extrusion process.

In the embodiment that is more preferably, the astaxanthin-containing extract per os of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part directly is applied to animal.

Before using, it is favourable that the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part is processed into a kind of form that can directly be administered orally in animal and preferably produce high stability and high bioavilability astaxanthin in use field separately.

Depend on the animal that it is carried out dosage forms for oral administration, and also depend on the animal feed goods that therefore multiple procedure of processing all can be favourable.

For the plants of tagetes species or the plant part of astaxanthin-containing, dry in this embodiment and/or pulverize the plant of astaxanthin-containing or plant part especially capitulum or petal are favourable.Particularly preferably, the plants of tagetes species of astaxanthin-containing or plant part exist with powder type.

Yet no matter the plants of tagetes species of the astaxanthin-containing that each embodiment is prepared or plant part are processed or are not processed, and all can be administered orally in animal in a manner known way.

If still the solvent of Cun Zaiing is harmless on the physiology for corresponding animal, the astaxanthin-containing extract can directly be administered orally in animal.

Evaporate after the solvent that still exists, extract can be applied with the form of astaxanthin-containing powder or oil.

For example, resulting astaxanthin-containing powder or oil can mix fish oil, can be applied to for example wheat flour or wipe broken marigold petal of dust carrier material, perhaps are included in alginates, gelatin or the lipid.

Therefore preferably liquid or powder type of astaxanthin-containing extract or processing back extract.

Yet no matter the astaxanthin-containing extract of the plants of tagetes species of the astaxanthin-containing that each embodiment is prepared or plant part is processed or is not processed, and all can be administered orally in animal in a manner known way.

Therefore the invention still further relates to the colouring agent of the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species that contains astaxanthin-containing or plant part or astaxanthin-containing or plant part, the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part as required can be as above-mentioned processed in this case.

In preferred embodiments, colouring agent contains plants of tagetes species or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part of astaxanthin-containing, and the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part as required can be as above-mentioned processed in this case.

In particularly preferred colouring agent, employed plant part is capitulum or petal.

The invention further relates to and be administered orally in animal by astaxanthin-containing extract and carry out animal or animal product method of colouring the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part.

The invention further relates to and be administered orally in animal by astaxanthin-containing extract and produce the painted animal or the method for animal product the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part.

The invention further relates to the purposes of the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part as animal feed or animal feed additive.

Contain the colouring agent of astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part or the advantage that the animal feed that contains these colouring agents also has pigment astaxanthin high storage stability and high bioavilability.

To describe the present invention by following examples now, but be not limited thereto:

Example I

Produce the genetically modified plant of Tagetes of astaxanthin-containing

The normal experiment condition:

The sequence analysis of recombinant DNA

(Sanger etc., Proc.Natl.Acad.Sci.USA 74 (1977), 5463-5467) use the laser fluorescence DNA-sequenator from Licor (to distribute by MWG Biotech, Ebersbach) recombinant DNA molecules is checked order by the Sanger method.

Example I .1:

Amplification coding is from the cDNA of the total length primary sequence of the ketolase of haematococcus pluvialis Flotow em.Wille

Increase from the cDNA of haematococcus pluvialis coding ketolase from haematococcus pluvialis (192.80 strains at G_ttingen university algal cultures preservation center) suspension culture by PCR method.

For from room temperature (1.2g/l sodium acetate, 2g/l yeast extract, 0.2g/l MgCl the haematococcus culture medium ₂6H ₂O, 0.02CaCl ₂2H ₂O; PH 6.8; Behind the autoclaving, add 400mg/lL-asparagine, 10mg/l FeSO ₄H ₂O) the indirect daylight total RNA of preparation in haematococcus pluvialis (192.80 strain) suspension culture in 2 weeks that grows, step is a harvesting, freezingly grinds in liquid nitrogen and in mortar.Be transferred to the alga cells of the freezing pulverizing of 100mg in the reaction vessel then and add in the 0.8ml Trizol buffer solution (LifeTechnologies).With 0.2ml chloroform extracting suspension.Behind centrifugal 15 minutes of the 12000g, take out moisture supernatant and it is transferred in the new reaction vessel and carries out extracting with the long-pending ethanol of monoploid.With the long-pending isopropanol precipitating RNA of monoploid, be dissolved in the DEPC water (room temperature will contain the water overnight incubation of 1/1000 volume pyrocarbonic acid diethyl ester, autoclaving then) with the washing of 75% ethanol and with sediment.By spectrophotometry RNA concentration.

Synthetic for cDNA, with 60 ℃ of sex change of the total RNA of 2.5 μ g 10 minutes, cooled on ice 2 minutes is also utilized cDNA kit (Ready-to-go-you-prime-beads, Pharmacia Biotech), use antisense-special primer (PR1 SEQ ID NO:29) that it is transcribed into cDNA according to the explanation of manufacturer.

Use one to have a mind to-special primer (PR2 SEQ ID NO:30) and an antisense-special primer (PR1 SEQ ID NO:29), increase from the nucleic acid of haematococcus pluvialis (192.80 strain) coding ketolase from haematococcus pluvialis by PCR (PCR).

The PCR condition is as follows:

The cDNA of the ketolase protein that pcr amplification coding is made up of the total length primary sequence in containing the 50ml reactant mixture of following composition:

-4ml haematococcus pluvialis cDNA (as above-mentioned preparation)

-0.25mM?dNTPs

-0.2mM?PR1(SEQ?ID?NO：29)

-0.2mM?PR2(SEQ?ID?NO：30)

-5ml 10 * PCR buffer solution (TAKARA)

-0.25ml R Taq polymerase (TAKARA)

-25.8ml distilled water.

PCR carries out under following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

53 ℃ 2 minutes

72 ℃ 3 minutes

1 * 72 ℃ 10 minutes

Use the 1155bp fragment (SEQ ID NO:22) of the pcr amplification generation coding total length primary sequence protein of SEQ ID NO:29 and SEQ ID NO:30.Use standard method, amplicons cloned is gone into PCR cloning vector pGEM-Teasy (Promega) and obtained cloning pGKETO2.

Sequencing analysis to the clone pGKETO2 that contains T7 and SP6 primer confirms that this sequence only has three codons 73,114 and 119 different with disclosed sequence X 86782, wherein has a base difference in each codon.In different amplification experiment, also produced these nucleotide substitutions and therefore represented nucleotide sequence (Fig. 1 and 2, sequence relatively) in employed haematococcus pluvialis 192.80 strains.

Therefore this clone is used to be cloned into expression vector pJIT117 (Guerineau etc., 1988, Nucl.Acids Res.16:11380).Clone by separating 1027bp SpHI fragment from pGEM-Teasy and connecting into the carrier pJIT117 that cuts through the SpHI enzyme.The clone who contains haematococcus pluvialis ketolase and N end and rbcs transit peptides translation fusion with correct direction is called pJKETO2.

Example I .2:

Amplification coding is from the terminal cDNA that shortens 14 amino acid whose ketolases of the N of haematococcus pluvialis Flotow em.Wille

Utilize the PCR method from the terminal cDNA that shortens 14 amino acid whose ketolases of haematococcus pluvialis suspension culture (192.80 strains at G_ttingen university algal cultures preservation center) amplification coding haematococcus pluvialis (192.80 strain) N.

As described in embodiment 1, prepare total RNA from haematococcus pluvialis (192.80 strain) suspension culture.

Synthetic as carrying out cDNA as described in the embodiment 1.

Utilize PCR (PCR) to use one to have a mind to special primer (PR3 SEQ IDNO:31) and an antisense special primer (PR1 SEQ ID NO:29) from the nucleic acid of haematococcus pluvialis amplification coding from 14 amino acid whose ketolases of the terminal shortening of the N of haematococcus pluvialis (192.80 strain).

The PCR condition is as follows:

The terminal cDNA that shortens 14 amino acid whose ketolase protein of pcr amplification coding N in containing the 50ml reactant mixture of following composition:

-4ml haematococcus pluvialis cDNA (as above-mentioned preparation)

-0.25mM?dNTPs

-0.2mM?PR1(SEQ?ID?NO：29)

-0.2mM?PR3(SEQ?ID?NO：31)

-5ml 10 * PCR buffer solution (TAKARA)

-0.25ml R Taq polymerase (TAKARA)

-25.8ml distilled water.

PCR carries out under following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

53 ℃ 2 minutes

72 ℃ 3 minutes

1 * 72 ℃ 10 minutes

Use the pcr amplification reaction of SEQ ID NO:29 and SEQ ID NO:31 to produce the 1111bp fragment of coding-terminal amino acid (position 2-16) by the ketolase protein of single amino acid (leucine) replacement.

Use standard method, amplicons cloned is gone into PCR cloning vector pGEM-Teasy (Promega).Use primer T7 to carry out sequencing with SP6 and confirm that this sequence is identical with sequence SEQ IDNO:22, difference is that the 5 ' zone (position 1-53) of SEQ ID NO:22 is replaced by nine different base sequences among the amplicon SEQ ID NO:24.Therefore this clone is used to be cloned into expression vector pJIT117 (Guerineau etc. 1988, Nucl.Acids Res.16:11380).

Clone by separating 985bp SpHI fragment from pGEM-Teasy and connecting into the carrier pJIT117 that cuts through the SpHI enzyme.Contain the terminal clone who shortens 14 amino acid whose haematococcus pluvialis ketolases and N-end and rbcs transit peptides fusion translation of N with correct direction and be called pJKETO3.

Example I .3:

(algal cultures preservation center 192.80 strains of G_ttingen university) contain the ketolase of whole primary sequences and merge the cDNA that the terminal myc-label of C-is arranged amplification coding from haematococcus pluvialis Flotow em.Wille

(192.80 strain) contains the ketolase of whole primary sequences and merges the cDNA that the terminal myc-label of C-is arranged from haematococcus pluvialis to utilize plasmid pGKETO2 (as described in example 1 above) and primer PR15 (SEQ ID NO:32) preparation coding by the PCR method.Primer PR15 comprises 5 ' zone (nucleotides 1-39) of special 3 ' zone (nucleotides 40-59) of antisense and coding myc-label.

In containing the 11.5ml reactant mixture of following composition, carry out sex change (95 ℃ 5 minutes) and the annealing (slowly cooling to 40 ℃) of pGKETO2 and PR15 in room temperature:

-1mg pGKETO2 DNA

-0.1mg?PR15(SEQ?ID?NO：32)

In containing the 20ml reactant mixture of following composition, mend flat 3 ' terminal (30 ℃ 30 minutes):

-11.5ml pGKETO2/PR15 annealing reaction liquid (producing as mentioned above)

-50mM?dNTPs

-2ml 1 * Ke Lienuo buffer solution

-2U Klenow enzyme

Use one to have a mind to special primer (PR2 SEQ IDNO:30) and an antisense special primer (PR15 SEQ ID NO:32) (192.80 strain) contains the ketolase of whole primary sequences and merge the nucleic acid that C end myc-label is arranged from haematococcus pluvialis from the haematococcus pluvialis amplification coding by PCR (PCR).

The PCR condition is as follows:

The pcr amplification coding has the cDNA of the ketolase protein that merges the terminal myc-label of C in containing the 50ml reaction mixture of following composition:

-1ml annealing reaction liquid (producing as mentioned above)

-0.25mM?dNTPs

-0.2mM?PR15(SEQ?ID?NO：32)

-0.2mM?PR2(SEQ?ID?NO：30)

-5ml 10 * PCR buffer solution (TAKARA)

-0.25ml R Taq polymerase (TAKARA)

-28.8ml distilled water.

Carry out PCR at following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

53 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

Use SEQ ID NO:32 and SEQ ID NO:30 pcr amplification to produce and encode and contain from whole primary sequences of haematococcus pluvialis ketolase and 1032bp fragment terminal at N and the fusion of rbcS transit peptides and and the dual translation fused protein that the myc-label merges terminal at C.

Use standard method, amplicons cloned is gone into PCR cloning vector pGEM-Teasy (Promega).Use primer T7 to carry out sequencing with SP6 and confirm that this sequence is identical with sequence SEQ IDNO:22, difference is replaced by different 39bp sequences by the 3 ' zone (position 993-1155) of SEQ ID NO:22 in amplicon SEQ ID NO:26.Therefore this clone is used to be cloned into expression vector pJIT117 (Guerineau etc. 1988, Nucl.Acids Res.16:11380).

Clone by separating 1038bp EcoRI-SpHI fragment from pGEM-Teasy and connecting into the carrier pJIT117 that cuts through the EcoRI-SpHI enzyme.Connect and cause between the C of rbcS transit peptide sequence end and ketolase sequence of N end, producing the translation fusion.Contain with correct direction and be called pJKETO4 the terminal clone of merging myc-label and and the haematococcus pluvialis ketolase that the translation of rbcS transit peptides is merged terminal of C at N.

Example I .4:

Amplification coding is from the DNA of the whole primary sequences of ketolase of beads algae (Nostoc sp.) PCC 7120

By the bacterial strain of PCR from beads algae PCC 7120 (" Pasteur cyanobacteria culture collection center (Pasteur Culture Collection of Cyanobacterium ")) amplification coding is from the DNA of beads algae PCC 7120 ketolases.

For from 25 ℃ in BG 11 culture mediums (1.5g/l NaNO ₃, 0.04g/lK ₂PO ₄3H ₂O, 0.075g/l MgSO ₄H ₂O, 0.036g/l CaCl ₂2H ₂O, 0.006g/l citric acid, 0.006g/l ferric citrate, 0.001g/l EDTA disodium magnesium, 0.04g/l Na ₂CO ₃, 1ml trace meter mixture A5+Co (2.86g/l H ₃BO ₃, 1.81g/l MnCl ₂4H ₂O, 0.222g/lZnSO ₄7H ₂O, 0.39g/l NaMoO ₄2H ₂O, 0.079g/l CuSO ₄5H ₂O, 0.0494g/lCo (NO ₃) ₂6H ₂O) prepare genomic DNA in beads algae PCC 7120 suspension cultures in continuous light and persistent oscillation (150 rev/mins) one week of growth in,, freezingly pulverize in liquid nitrogen and in mortar by centrifugal cell harvesting.

Step from beads algae PCC 7120 DNA isolations:

By 8000 rev/mins centrifugal 10 minutes, from the 10ml liquid culture, be settled out bacterial cell.Use mortar in liquid nitrogen, to push and grind bacterial cell then.Cellular material is resuspended in 1ml 10mM Tris HCl (pH 7.5) and is transferred in the Eppendorf reaction vessel (2ml volume).Add 100 μ l Proteinase Ks (concentration: 20 mg/ml), cell suspension was hatched 3 hours at 37 ℃.Use 500 μ l phenol extracting suspension then.13000 rev/mins after centrifugal 5 minutes, the water on upper strata is transferred in the new 2ml Eppendorf reaction tube.Repeat phenol extracting 3 times.Add the 3 M sodium acetates (pH 5.2) of 1/10 volume and the isopropanol precipitating DNA of 0.6 volume, wash with 70% ethanol then.Drying at room temperature DNA precipitation adds 25 μ l water then 65 ℃ of heating for dissolving.

By PCR (PCR), use one to have a mind to special primer (NOSTF, SEQID No.87) and an antisense special primer (NOSTG, SEQ ID NO.88) from the nucleic acid of beads algae PCC7120 amplification coding from the ketolase of beads algae PCC 7120.

The PCR condition is as follows:

The pcr amplification coding contains the ketolase protein DNA of whole primary sequences in containing 50 μ l reaction mixtures of following composition:

-1 μ l beads algae PCC 7120 DNA (preparation as mentioned above)

-0.25mM?dNTPs

-0.2mM?NOSTF(SEQ?ID?No.87)

-0.2mM?NOSTG(SEQ?ID?No.88)

-5 μ l 10 * PCR buffer solutions (TAKARA)

-0.25 μ l R Taq polymerase (TAKARA)

-25.8 μ l distilled water.

Under following cycling condition, carry out PCR:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

55 ℃ 1 minute

72 ℃ 3 minutes

1 * 72 ℃ 10 minutes

Use SEQ ID NO:87 and SEQ ID NO:88 pcr amplification to produce and encode and contain the 805bp fragment (SEQ ID NO:89) of the whole primary sequences of protein.Use standard method, amplicons cloned is gone into PCR cloning vector pGEM-T (Promega) and obtained cloning pNOSTF-G.

Use M13F to check order at clone pNOSTF-G and confirm that this sequence is identical with the dna sequence dna of database typing AP003592 with the M13R primer.In different amplification experiments, also produced this nucleotide sequence, so it has represented the nucleotide sequence among the employed beads algae PCC 7120.

Therefore cloning pNOSTF-G is used to be cloned into expression vector pJIT117 (Guerineau etc. 1988, Nucl.Acids Res.16:11380).Clone by separating 1027bp SpHI fragment from pGEM-T and connecting into the carrier pJIT117 that cuts through the SpHI enzyme.Contain the Nostoc ketolase and be called pJNOST with correct direction the terminal clone of merging of its N with the translation of rbcS transit peptides.

Example I .5:

Generation is used for the expression vector at marigold constitutive expression haematococcus pluvialis ketolase.

Ketolase from haematococcus pluvialis is expressed (Franck etc. 1980, Cell 21:285-294) in marigold under the constitutive promoter d35S of CaMV control.Use is expressed (Anderson etc. 1986, and Biochem J.240:709-715) from the transit peptides rbcS of pea.

Produce the expression cassette be used for carrying out agriculture bacillus mediated conversion from the haematococcus pluvialis ketolase marigold by binary vector pSUN5 (WO 02/00900).

In order to produce marigold expression vector pS5KETO2, will be connected (Fig. 3, construct figure) with the carrier pSUN5 that cuts through the SacI-XhoI enzyme from the 2.8KbSacI-XhoI fragment of pJKETO2.In Fig. 3,35S promoter (747bp), fragment rbcS that fragment d35s contains repetition are that the rbcS transit peptides (204bp) from pea, whole primary sequences (1027bp), the fragment term of fragment KETO2 coding haematococcus pluvialis ketolase are CaMV polyadenylation signal (761bp).

Example I .5A:

Generation is used for the expression vector at marigold specifically expressing haematococcus pluvialis ketolase

Use is expressed ketolase (Anderson etc. 1986, and Biochem J.240:709-715) from haematococcus pluvialis from the transit peptides rbcS of pea in marigold.At arabidopsis cauliflower specific promoter (AL132971: nucleotides zone 9298-10 200; Hill etc. (1998) Development 125:1711-1721) expresses under the control of modified forms AP3P.

Utilize genomic DNA (separating from mouseearcress) and primer PR7 (SEQ ID NO:33) and PR10 (SEQ ID NO:36) to produce the dna fragmentation that contains mouseearcress AP3 promoter region-902-+15 by PCR by standard method.

The PCR condition is as follows:

In containing the 50ml reaction mixture of following composition pcr amplification contain the AP3 promoter fragment (DNA 902-+15):

-100ng mouseearcress genomic DNA

-0.25mM?dNTPs

-0.2mM?PR7(SEQ?ID?NO：33)

-0.2mM?PR10(SEQ?ID?NO：36)

-5ml 10 * PCR buffer solution (Stratagene)

-0.25ml Pfu polymerase (Stratagene)

-28.8ml distilled water.

PCR is undertaken by following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

50 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

Use standard method that the 922bp amplicons cloned is gone into PCR cloning vector pCR 2.1 (Invitrogen), and obtain plasmid pTAP3.

Sequencing to clone pTAP3 confirms that its sequence and disclosed AP3 sequence (AL132971, nucleotides zone 9298-10200) only exist an insertion (9765 of sequence A L132971 are inserted G) to replace the different of (9726 the A of sequence A L132971 replaces with G) with a base.The difference that these nucleotides in different amplification experiments, also occurred, so this sequence has been represented nucleotide sequence actual in the used mouseearcress plant.

Utilize plasmid pTAP3 to produce the AP3P of modified forms by the recombinant PCR method.Use primer PR7 (SEQ ID NO:33) and PR9 (SEQ ID NO:35) amplification region 10200-9771 (amplicon A7/9), use PR8 (SEQ ID NO:34) and PR10 (SEQ ID NO:36) amplification region 9526-9285 (amplicon A8/10).

The PCR condition is as follows:

Pcr amplification contains the dna fragmentation of AP3 promoter region 10200-9771 and regional 9526-9285 in the 50ml reactant mixture, and wherein composition is as follows:

-100ng AP3 amplicon (as mentioned above)

-0.25mM?dNTP

-0.2mM has a mind to primer (PR7 SEQ ID NO:33 or PR8 SEQ ID NO:34)

-0.2mM antisense primer (PR9 SEQ ID NO:35 or PR10 SEQ ID NO:36)

-5ml 10 * PCR buffer solution (Stratagene)

-0.25ml Pfu Taq polymerase (Stratagene)

-28.8ml distilled water.

PCR is undertaken by following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

50 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

The recombinant PCR reaction comprises anneals to 25 overlapping amplicon A7/9 and A8/10 of nucleotide sequence, forms double-stranded and amplification subsequently.This produces the modified forms AP3P of AP3 promoter, wherein 9670-9526 position disappearance.In containing the 17.6ml reactant mixture of following composition, carry out sex change (95 ℃ 5 minutes) and the annealing (room temperature slowly cools to 40 ℃) of two amplicon A7/9 and A8/10:

-0.5mg A7/9 amplicon

-0.25mg A8/10 amplicon

In containing the 20ml reactant mixture of following composition, mend flat (30 ℃ 30 minutes) 3 ' end:

-17.6ml gA7/9 and A8/10 annealing reaction liquid (producing as mentioned above)

-50mM?dNTP

-2ml 1 * Ke Lienuo buffer solution

-2U Klenow enzyme

Use a nucleic acid of having a mind to special primer (PR7 SEQ ID NO:33) and an antisense special primer (PR10 SEQ ID NO:36) the modified form AP3P of amplification coding promoter by PCR method.

The PCR condition is as follows:

Pcr amplification AP3P fragment in containing the 50ml reactant mixture of following composition:

-1ml annealing reaction liquid (producing as mentioned above)

-0.25mM?dNTP

-0.2mM?PR7(SEQ?ID?NO：33)

-0.2mM?PR10(SEQ?ID?NO：36)

-5ml 10 * PCR buffer solution (Stratagene)

-0.25ml Pfu Taq polymerase (Stratagene)

-28.8ml distilled water.

PCR is undertaken by following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

50 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

The pcr amplification that uses SEQ ID NO:33 and SEQ ID NO:36 to carry out produces the 778bp fragment of the modified form AP3P of coding promoter.Amplicons cloned is gone into cloning vector pCR2.1 (Invitrogen).Use the sequencing of primer T7 and M13 to confirm that its sequence is identical with the regional 10200-9298 that sequence A L132971 interior zone 9285-9526 has lacked.Therefore this clone is used to be cloned into expression vector pJIT117 (Guerineau etc. 1988, Nucl.Acids Res.16:11380).

Clone by separating 771bp SacI-HindIII fragment from pTAP3P and connecting into the carrier pJIT117 that cuts through the SacI-HindIII-enzyme.The clone who contains promoter AP3P rather than original promoter d35S is called pJAP3P.

In order to produce expression cassette pJAP3PKETO2, KETO2 is cloned in the carrier pJAP3P that the SpHI-enzyme is cut with 1027bp SpHI fragment.The fragment KETO2 and the terminal clone of merging with the rbcS transit peptides of its N-that contain correct direction are called pJAP3PKETO2.

In order to produce expression cassette pJAP3PKETO4,1032bp SpHI-EcoRI fragment KETO4 (being described in embodiment 3) is cloned in the carrier pJAP3P that the SpHI-EcoRI-enzyme is cut.The fragment KETO4 and the terminal clone of merging with the rbcS transit peptides of its N-that contain correct direction are called pJAP3PKETO4.

Use binary vector pSUN5 (WO 02/00900) preparation to be used for carrying out the expression vector that agriculture bacillus mediated haematococcus pluvialis is subjected to the conversion of AP3P-control ketolase marigold.

In order to produce expression vector pS5AP3PKETO2, will connect into the carrier pSUN5 (Fig. 4, construct figure) that cuts through the SacI-XhoI-enzyme from the 2.8KB bp SacI-XhoI fragment of pJAP3PKETO2.In Fig. 4, Segment A P3P contains modified AP3P promoter (771bp), fragment rbcS contains the rbcS transit peptides (204bp) of pea, fragment KETO2 (1027bp) contains whole primary sequences of coding haematococcus pluvialis ketolase, and fragment term (761bp) contains the CaMV polyadenylation signal.

Example I .5.B:

Generation is used for reading at the marigold constitutive expression expression vector of ball algae PCC 7120 ketolases

The Nostoc ketolase is expressed under mouseearcress constitutive promoter FNR (ferredoxin-NADPH oxidoreducing enzyme) control in the marigold.Use pea transit peptides rbcS to express (Anderson etc. 1986, and Biochem J.240:709-715).

Utilize genomic DNA (using standard method to separate) and primers F NR-1 (SEQ ID No.90) and FNR-2 (SEQ ID No.91) to produce the dna fragmentation that contains mouseearcress FNR promoter region-635--1 by PCR method from mouseearcress.

The PCR condition is as follows:

In containing 50 μ l reactant mixtures of following composition pcr amplification contain FNR promoter fragment FNR1-2 (DNA 635--1):

-100ng mouseearcress genomic DNA

-0.25mM?dNTP

-0.2mM?FNR-1(SEQ?ID?No.90)

-0.2mM?FNR-2(SEQ?ID?No.91)

-5 μ l 10 * PCR buffer solutions (Stratagene)

-0.25 μ l Pfu polymerase (Stratagene)

-28.8 μ l distilled water.

PCR carries out according to following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

50 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

By standard method the 653bp amplicons cloned is gone into PCR cloning vector pCR 2.1 (Invitrogen) and obtained plasmid pFNR.

Clone pFNR is carried out sequencing confirm that No. 5 chromosome of its sequence and mouseearcress is from 70127-69493 bit sequence part consistent (data base access AB011474).This gene originates in base-pair 69492 and is called " ferredoxin-NADP ⁺Reductase ".

This clone is called pFNR and is used to be cloned into expression vector pJIT117 (Guerineau etc. 1988, Nucl.Acids Res.16:11380).

Clone by separating 635bp SacI-HindIII fragment from pFNR and connecting into the carrier pJIT117 that cuts through the SacI-HindIII-enzyme.The clone who contains promoter FNR rather than original promoter d35S is called pJITFNR.

In order to produce expression cassette pJFNRNOST, 805bp SpHI fragment NOSTF-G (being described in embodiment 1) is cloned into the carrier pJITFNR that cuts through the SpHI-enzyme.The fragment NOSTF-G and the terminal clone of merging with the rbcS transit peptides of its N-that contain correct direction are called pJFNRNOST.

Use binary vector pSUN5 (WO 02/00900) to produce and be used for carrying out the expression cassette that the agriculture bacillus mediated expression vector with Nostoc ketolase transforms marigold.

In order to produce marigold expression vector pS5FNRNOST, the 2.4KbSacI-XhoI fragment (part SacI hydrolysis) of pJFNRNOST is connected into the carrier pSUN5 (Fig. 5, construct figure) that cuts through the SacI-XhoI-enzyme.In Fig. 5, fragment FNR promoter contains the FNR promoter (655bp) of repetition, fragment rbcS transit peptides contains pea rbcS transit peptides (204bp), fragment Nost ketolase (799bp) contains whole primary sequences of coding Nostoc ketolase, and fragment 35S Term (761bp) contains the CaMV polyadenylation signal.

Example I .5C:

Generation is used for the expression vector at marigold specifically expressing beads algae PCC 7120 ketolases

Use pea transit peptides rbcS in marigold, to express Nostoc ketolase (Anderson etc. 1986, and Biochem J.240:709-715).Be expressed in arabidopsis cauliflower specific promoter AP3 (AL132971: nucleotides zone 9298-10200; Hill etc. (1998) Development 125:1711-1721) carries out under the modified form AP3P control.

Utilize genomic DNA (using standard method to separate) and primer AP3-1 (SEQ ID No.93) and AP3-2 (SEQ ID No.94) to produce the dna fragmentation that contains mouseearcress AP3 promoter region-902-+15 by PCR from mouseearcress.

The PCR condition is as follows:

In containing 50 μ l reactant mixtures of following composition pcr amplification contain the AP3 promoter fragment (DNA 902-+15):

-100ng mouseearcress genomic DNA

-0.25mM?dNTP

-0.2mM?AP3-1(SEQ?ID?No.93)

-0.2mM?AP3-2(SEQ?ID?No.94)

-5 μ l 10 * PCR buffer solutions (Stratagene)

-0.25 μ l Pfu polymerase (Stratagene)

-28.8 μ l distilled water.

PCR carries out according to following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

50 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

Use standard method that the 929bp amplicons cloned is gone into PCR cloning vector pCR 2.1 (Invitrogen) and obtained plasmid pAP3.

Clone pAP3 is carried out sequencing confirm that this sequence and disclosed AP3 sequence (AL132971, nucleotides zone 9298-10200) only exist an insertion (9765 of sequence A L132971 are inserted a G) to replace the different of (9726 the A of sequence A L132971 replaces with G) with a base.In different amplification experiments, also produced these nucleotides differences, so this sequence has been represented nucleotide sequence actual in the employed mouseearcress plant.

Utilize plasmid pAP3 to produce modified forms AP3P by the recombinant PCR method.Use primer AP3-1 (SEQ ID No.93) and AP3-4 (SEQ ID No.96) amplification region 10200-9771 (amplicon A1/4), use AP3-3 (SEQ ID No.95) and AP3-2 (SEQ ID No.94) amplification region 9526-9285 (amplicon A2/3).

The PCR condition is as follows:

Pcr amplification contains the dna fragmentation of AP3 promoter region 10200-9771 and regional 9526-9285 in containing 50 μ l reactant mixtures of following composition:

-100ng AP3 amplicon (as mentioned above)

-0.25mM?dNTPs

-0.2mM has a mind to primer (AP3-1 SEQ ID No.93 or AP3-3 SEQ ID No.95)

-0.2mM antisense primer (AP3-4 SEQ ID No.96 or AP3-2 SEQ ID No.94)

-5 μ l 10 * PCR buffer solutions (Stratagene)

-0.25 μ l Pfu Taq polymerase (Stratagene)

-28.8 μ l distilled water.

PCR carries out according to following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

50 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

Recombinant PCR comprises that there are 25 amplicon A1/4 and A2/3 that nucleotide sequence is overlapping in annealing, forms double-stranded and amplification subsequently.This produces the modified form AP3P of AP3 promoter, wherein 9670-9526 position disappearance.In containing 17.6 μ l reactant mixtures of following composition, carry out sex change (95 ℃ 5 minutes) and the annealing (room temperature slowly cools to 40 ℃) of two amplicon A1/4 and A2/3:

-0.5 μ g A1/4 amplicon

-0.25 μ g A2/3 amplicon

In containing 20 μ l reactant mixtures of following composition, mend flat (30 ℃ 30 minutes) 3 ' end:

-17.6 μ l A1/4 and A2/3 annealing reaction liquid (producing as mentioned above)

-50μM?dNTPs

-2 μ l 1 * Ke Lienuo buffer solution

-2U Klenow enzyme

Use a nucleic acid of having a mind to special primer (AP3-1 SEQ ID No.93) and an antisense special primer (AP3-2 SEQ ID No.94) the modified form AP3P of amplification coding promoter by PCR method.

The PCR condition is as follows:

Pcr amplification AP3P fragment in containing 50 μ l reactant mixtures of following composition:

-1 μ 1 annealing reaction liquid (producing as mentioned above)

-0.25mM?dNTPs

-0.2mM?AP3-1(SEQ?ID?No.93)

-0.2mM?AP3-2(SEQ?ID?No.94)

-5 μ l 10 * PCR buffer solutions (Stratagene)

-0.25 μ l Pfu Taq polymerase (Stratagene)

-28.8 μ l distilled water.

PCR carries out according to following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

50 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

Use SEQ ID No.93 (AP3-1) and SEQ ID No.94 (AP3-2) to carry out the 783bp fragment that pcr amplification produces the modified form AP3P of coding promoter.Amplicons cloned is gone into cloning vector pCR2.1 (Invitrogen) and obtained plasmid pAP3P.Use primer T7 to carry out sequencing and confirm that this sequence is identical with the regional 10200-9298 that sequence A L132971 interior zone 9285-9526 has lacked with M13.Therefore this clone is used to be cloned into expression vector pJIT117 (Guerineau etc. 1988, Nucl. Acids Res.16:11380).

Clone by separating 783bp SacI-HindIII fragment from pAP3P and connecting into the carrier pJIT117 that cuts through the SacI-HindIII-enzyme.The clone who contains promoter AP3P rather than original promoter d35S is called pJITAP3P.In order to produce expression cassette pJAP3NOST, 805bp SpHI fragment NOSTF-G (being described in embodiment 1) is cloned into the carrier pJITAP3P that cuts through the SpHI-enzyme.The fragment NOSTF-G and the terminal clone of merging with the rbcS transit peptides of its N-that contain correct direction are called pJAP3PNOST.

Use binary vector pSUN5 (WO 02/00900) generation to be used for carrying out the expression cassette of the Nostoc ketolase conversion of agriculture bacillus mediated AP3P control marigold.

In order to produce expression vector pS5AP3PNOST, the 2.6KbSacI-XhoI fragment (part SacI hydrolysis) of pS5AP3PNOST is connected into the carrier pSUN5 (Fig. 6, construct figure) that cuts through the SacI-XhoI-enzyme.In Fig. 6, Segment A P3P contains modified AP3P promoter (783bp), fragment rbcS contains pea rbcS transit peptides (207bp), and fragment NOSTF-G (792bp) contains whole primary sequences of coding Nostoc ketolase, and fragment term (795bp) contains the CaMV polyadenylation signal.

Example I .6:

The generation of transgenosis marigold plant

With marigold seed sterilization and be positioned over germination culture medium (MS culture medium; Murashige and Skoog, Physiol.Plant.15 (1962), 473-497) pH 5.8,2% sucrose).Germination is to carry out in 18-28 ℃/20-200mE/3-16 week in temperature/illumination/time interval, is preferably 21 ℃, and 20-70mE cultivates 4-8 week.

Gather in the crops all ectogenetic plant leafs and transverse cuts till that time to middle arteries and veins.In preparation process, be 10-60mm with the size that is produced ²The leaf explant room temperature preservation the longest 2 hours in liquid MS medium.

Optional Agrobacterium tumefaciems bacterial strain but be preferably highly virulent strain and for example have the EHA105 grow overnight of the corresponding double base plasmid (for example pS5KETO2 and pS5AP3PKETO2) that carries selectable marker gene (preferably bar or pat) and one or more characteristic or reporter gene and be used for cultivating altogether with the leaf material.The growth of bacterial isolates is following to be carried out: the monospecific polyclonal of corresponding bacterial strain is inoculated in YEB (0.1% yeast extract, 0.5% beef extract, 0.5% peptone, 0.5% sucrose, the 0.5% magnesium sulfate 7H that contains the 25mg/l kanamycins ₂O) cultivated 16-20 hour in and at 28 ℃.Pass through then 6000g centrifugal 10 minutes results bacterial suspension and with every OD ₆₀₀Approximately the concentration of 0.1-0.8 is resuspended in the liquid MS medium.This suspension is used for cultivating altogether with the leaf material.

Before cultivating altogether, replace preserving the MS culture medium of leaf immediately with bacterial suspension.The room temperature of vibrating was gently hatched leaf in the Agrobacterium suspension 30 minutes.Then the explant that infects being positioned over agar solidifies and (contains growth regulator for example on the MS culture medium of 3mg/l benayl aminopurine (BAP) and 1mg/l heteroauxin (IAA) as 0.8% plant agar (Duchefa, NL)).The location of leaf on culture medium is not important.Explant is cultivated 1-8 days, but be preferably 6 days, can use following condition: luminous intensity: 30-80mmol/m ²X second, temperature: 22-24 ℃, light/secretly be replaced by 16/8 hour.Then, the explant of cultivating altogether is transferred to fresh MS culture medium, is preferably the fresh MS culture medium that contains the isometric growth conditioning agent, this second culture medium additionally contains the antibiotic of bacteria growing inhibiting.Concentration is that the Ticarcillin/Clavulanate Acid (Timentin) of 200-500mg/l is particularly suitable for this purpose.As second selection component, use be to be used to screen successfully the composition that transforms.Be that the phosphinothricin of 1-5mg/l selects with concentration be very effective, but also can consider to use other selection component according to this method.

After each situation cultivated for one to three week, explant is transferred in the fresh culture up to plumule and budlet forms, then they are transferred to and contain the composition that Ticarcillin/Clavulanate Acid and PPT or other contain conditioning agent and just refer to such as 0.5mg/l indolebutyric acid (IBA) and 0.5mg/l gibberellic acid GA ₃The same basic culture medium in to take root.The branch of taking root can be transferred to the greenhouse.

Except said method, following promising change also is possible:

Before the bacterial infection explant, can with they in above-mentioned culture medium preincubate 1-12 days, be preferably 3-4 days, to be used for common cultivation.Then, infect as mentioned above, cultivate altogether and selectivity regeneration.

The pH that is used to regenerate (being generally 5.8) can be reduced to pH 5.2.This has improved the control to the Agrobacterium growth.

In regeneration culture medium, add AgNO ₃(3-10mg/l) improve cultivation conditions, comprised regeneration itself.

Reduction phenol forms and is for example citric acid, ascorbic acid, PVP and many other compositions of composition well known to those skilled in the art, and cultivation is had beneficial effect.

For all method, also can use fluid nutrient medium.Hatch on the conventional holder that culture also can the commerce in being placed on fluid nutrient medium can get.

According to above-mentioned method for transformation, use following expression construct to obtain following strain:

For example, use pS5KETO2 to obtain following strain: cs18-1 and cs18-2, for example use pS5AP3PKETO2 to obtain following strain: cs19-1, cs19-2 and cs19-3.For example, use pS5FNRNOST to obtain following strain: ms 103-1, ms103-2, ms103-3 for example, uses pS5AP3NOST to obtain following strain: ms 104-1, ms104-2, ms104-3.

Example I .8

The feature of genetically modified plants

Example I .8.1

The separation of carotenoid ester class in the genetically modified plants floral leaf

The overall work explanation:

In liquid nitrogen, grind the floral leaf of genetically modified plants also with 100% acetone extracting (3 times, each 500ml) petal powder (approximately 40mg) with mortar.Evaporating solvent and with carotenoid be resuspended in 100-200ml benzinum/acetone (5: 1, v/v).

According to their hydrophobicity, pass through at organic mobile phase (benzinum/acetone with the form that concentrates; 5: 1) in go up by TLC (TLC) in silica 60 F254 plates (Merck) and to separate carotenoid.On TLC, scrape yellow (lutein ester), red (keto-acid carotenoid ester) and orange (mixture of lutein ester and keto-acid carotenoid ester) band.

With the carotenoid of three wash-out silica bound of 500ml acetone, evaporating solvent also separates carotenoid by the HPLC method and also identifies.

By the C30 reversed-phase column, can region class carrotene monoesters and diester.The HPLC condition of work is in fact identical with disclosed method, and (Frazer etc. (2000), Plant Journal 24 (4): 551-558).Can identify carotenoid based on UV-VIS spectrum.

Example I .9

The enzyme hydrolysis of carotenoid ester and the evaluation of carotenoid

The overall work explanation

The petal material (50-100mg fresh weight) that in mortar, grinds with 100% acetone extracting (, vibrating about 15 minutes) at every turn with 500ml extracting 3 times.Evaporating solvent is dissolved in carotenoid 400ml acetone (475nm is absorbed between the 0.75-1.25) then and handled in ultra sonic bath 5 minutes.Carotenoid extract and 300ml 50mM Tris-HCl buffer solution (pH 7.0) mixed be incorporated in 37 ℃ and hatched 5-10 minute.Afterwards, add 100-200ml cholesterol esterase (storage liquid: 6.8 units/ml cholesterol esterase of pseudomonas (Pseudomonas spec.)).After 8-12 hour, add the 100-200ml enzyme again; In 24 hours, hatch the ester hydrolysis class at 37 ℃.Adding 0.35gNa ₂SO ₄10H ₂Behind O and the 500ml benzinum, mixture is fully mixed and centrifugal (3 minutes; 4500g).Take out benzinum mutually and again with 0.35g Na ₂SO ₄10H ₂O (anhydrous) mixes.Centrifugal 1 minute of 10000g.Evaporate benzinum and dissociation carrotene is dissolved in 100-120ml acetone.Use HPLC and C30 reversed-phase column method, can identify dissociation carrotene based on retention time and UV-VIS spectrum.

Example I .10:

Generation is used for the cloning vector at the reverse repetition expression cassette of marigold specifically expressing ε-cyclase ds RNA

In marigold in arabidopsis cauliflower specific promoter AP3 (AL132971: nucleotides zone 9298-10200; Hill etc. (1998) Development 125:1711-1721) the reverse repetition transcript of being made up of ε-cyclase fragment is expressed in modified form AP3P control down.

Under each situation, oppositely repeating transcript, to contain by functional introne be a potato ST-LH1 gene PIV2 introne forward fragment connected to one another (have a mind to fragment) and a reverse identical sequence fragment (antisense fragment) (1990) Mol Gen Genet220:245-50 such as () Vancanneyt G..

By PCR method, utilize genomic DNA (separating from mouseearcress) and primer PR7 (SEQ ID NO:49) and PR10 (SEQ ID NO:52) to produce coding mouseearcress AP3 promoter (cDNA 902-+15) by standard method.

The PCR condition is as follows:

Pcr amplification coding AP3 promoter fragment in containing the 50ml reactant mixture of following composition (DNA 902-+15):

-1ml mouseearcress genomic DNA (1: 100 dilution of Chan Shenging as mentioned above)

-0.25mM?dNTP

-0.2mM?PR7(SEQ?ID?NO：49)

-0.2mM?PR10(SEQ?ID?NO：52)

-5ml 10 * PCR buffer solution (Stratagene)

-0.25ml Pfu polymerase (Stratagene)

-28.8ml distilled water.

PCR carries out according to following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

50 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

Use standard method that the 922bp amplicons cloned is gone into PCR cloning vector pCR 2.1 (Invitrogen) and obtained plasmid pTAP3.Clone pTAP3 is carried out sequencing confirm this sequence and disclosed AP3 sequence (AL132971, nucleotides zone 9298-10200) only there is insertions (9765 of sequence A L132971 are inserted a G) alternative with a base (9726 the A of sequence A L132971 replaces with G) different (33: G replaces with T, and 55: G replaces with T).In different amplification experiments, also produce the difference of these nucleotides, therefore represented the nucleotide sequence in the employed mouseearcress.

Utilize plasmid pTAP3 to produce modified forms AP3P by the recombinant PCR method.Use primer PR7 (SEQ ID NO:49) and primer PR9 (SEQ ID NO:51) amplification 10200-9771 zone (amplicon A7/9), use PR8 (SEQ ID NO:50) and PR10 (SEQ ID NO:52) amplification 9526-9285 zone (amplicon A8/10).

The PCR condition is as follows:

The dna fragmentation in pcr amplification coding AP3 promoter 10200-9771 zone and 9526-9285 zone in containing the 50ml reactant mixture of following composition:

-100ng AP3 amplicon (as mentioned above)

-0.25mM?dNTPs

-0.2mM PR7 (SEQ ID NO:49) or PR8 (SEQ ID NO:50)

-0.2mM PR9 (SEQ ID NO:51) or PR10 (SEQ ID NO:52)

-5ml 10 * PCR buffer solution (Stratagene)

-0.25ml Pfu Taq polymerase (Stratagene)

-28.8ml distilled water.

PCR carries out according to following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

50 ℃ 2 minutes

72 ℃ 3 minutes

1 * 72 ℃ 10 minutes

The recombinant PCR method comprises having overlapping amplicon A7/9 of 25 nucleotide sequences and the annealing of A8/10, forms amplification double-stranded and subsequently.This has produced the modified forms AP3P of AP3 promoter, and wherein the 9670-9526 position is lacked.In containing the 17.6ml reactant mixture of following composition, carry out sex change (95 ℃ 5 minutes) and the annealing (room temperature slowly cools to 40 ℃) of two amplicon A7/9 and A8/10:

-0.5mg?A7/9

-0.25mg?A8/10

-17.6ml A7/9 and A8/10 annealing reaction liquid (producing as mentioned above)

-50mM?dNTPs

-2ml 1 * Ke Lienuo buffer solution

-2U Klenow enzyme

Use a nucleic acid of having a mind to special primer (PR7 SEQ ID NO:49) and an antisense special primer (PR10 SEQ ID NO:52) the modified form AP3P of amplification coding promoter by PCR method.

The PCR condition is as follows:

-1ml annealing reaction liquid (producing as mentioned above)

-0.25mM?dNTPs

-0.2mM?PR7(SEQ?ID?NO：49)

-0.2mM?PR10(SEQ?ID?NO：52)

-5ml 10 * PCR buffer solution (Stratagene)

-0.25ml Pfu Taq polymerase (Stratagene)

-28.8ml distilled water.

PCR carries out according to following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

50 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

Use PR7, the pcr amplification that SEQ ID NO:49 and PR10 SEQ ID NO:52 carry out produces the 778bp fragment of the modified form AP3P of coding promoter.Amplicons cloned is gone into cloning vector pCR2.1 (Invitrogen).Use the sequencing of primer T7 and M13 to confirm that this sequence is identical with the 10200-9298 zone that sequence A L132971 interior zone 9285-9526 has lacked.Therefore this clone is used to be cloned into expression vector pJIT117 (Guerineau etc. 1988, Nucl.Acids Res.16:11380).

Clone by separating 771bp SacI-HindIII fragment from pTAP3P and connecting into the carrier pJIT117 that cuts through the SacI-HindIII-enzyme.Contain promoter AP3P but not the clone of original promoter d35S is called pJAP3P.

Utilize DNA p35SGUS INT (1990) Mol Gen Genet 220:245-50 such as () Vancanneyt G and primer PR40 (Seq ID NO:54) and PR41 (SeqID NO:55) amplification to contain the dna fragmentation of gene ST-LS1 introne PIV2 by PCR method.

The PCR condition is as follows:

Pcr amplification gene ST-LS1 introne PIV2 sequence in containing the 50ml reactant mixture of following composition:

-1ml?p35SGUS?INT

-0.25mM?dNTPs

-0.2mM?PR40(SEQ?ID?NO：54)

-0.2mM?PR41(SEQ?ID?NO：55)

-5ml 10 * PCR buffer solution (TAKARA)

-0.25ml R Taq polymerase (TAKARA)

-28.8ml distilled water.

PCR carries out according to following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

53 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

The pcr amplification reaction that uses PR40 and PR41 to carry out produces the 206bp fragment.Use standard method, amplicons cloned is gone into PCR cloning vector pBluntII (Invitrogen) and obtained cloning pBluntII-40-41.Use primer SP6 that this clone is carried out sequencing and confirm that this sequence is with identical from the corresponding sequence of carrier p35SGUS INT.

Therefore this clone is used to be cloned into carrier pJAP3P (as mentioned above).

Clone by separating 206bp SaH-BamHI fragment from pBluntII-40-41 and connecting into the carrier pJAP3P that cuts through the SalI-BamHI-enzyme.Contain correct direction ST-LS1 gene PIV2 introne and be connected in the terminal clone of rbcs transit peptides 3 ' and be called pJAI1 and this clone and be suitable for producing and be used for the expression cassette that the flower specifically expressing oppositely repeats transcript.

In Fig. 7, fragment 4P3P contains that modified AP3P promoter (771bp), fragment rbcs contain pea rbcS transit peptides (204bp), fragment intron contains potato ST-LS1 gene PIV2 introne and fragment term (761bp) contains the CaMV polyadenylation signal.

Example I .11

Generation is used for the reverse repetition expression cassette (at the 5 ' zone of ε-cyclase cDNA) at marigold specifically expressing ε-cyclase dsRNA

Utilize one to have a mind to special primer (PR42 SEQ IDNO:56) and an antisense special primer (PR43 SEQ ID NO:57) and increase from marigold cDNA and contain ε-cyclase cDNA (Genbank accession number: the AF251016) nucleic acid in 5 '-terminal 435bp zone by PCR (PCR) method.Marigold ε-cyclase cDNA 5 '-terminal 435 bp zones comprise 138bp 5 '-non-translated sequence (5 ' UTR) and 297bp corresponding N end code area.

In order to prepare total RNA of marigold flower, the flower of the freezing pulverizing of 100mg is transferred to reaction vessel and is dissolved in 0.8ml Trizol buffer solution (LifeTechnologies).With 0.2ml chloroform extracting suspension.Behind centrifugal 15 minutes of the 12000g, take out water and it is transferred to new reaction tube and with the long-pending alcohol extract of monoploid.With the long-pending isopropanol precipitating RNA of monoploid, wash and precipitation is dissolved in DEPC water (room temperature with the pyrocarbonic acid diethyl ester night incubation of 1/1000 volume, autoclaving then) with 75% ethanol.By spectrophotometry RNA concentration.Synthetic for cDNA, with the total RNA of 2.5 μ g 60 ℃ of sex change 10 minutes, cooled on ice 2 minutes is also utilized cDNA kit (Ready-to-go-you-prime-beads, Pharmacia Biotech), use antisense special primer (PR17 SEQ ID NO:53) that it is transcribed into cDNA according to the operation instruction of manufacturer.

PCR reaction condition subsequently is as follows:

Pcr amplification contains the PR42-PR43 dna fragmentation in ε-cyclase 5 '-terminal 435bp zone in containing the 50ml reactant mixture of following composition:

-1ml cDNA (producing as mentioned above)

-0.25mM?dNTPs

-0.2mM?PR42(SEQ?ID?NO：56)

-0.2mM?PR43(SEQ?ID?NO：57)

-5ml 10 * PCR buffer solution (TAKARA)

-0.25ml R Taq polymerase (TAKARA)

-28.8ml distilled water..

Pcr amplification contains the PR44-PR45 dna fragmentation in ε-cyclase 5 '-terminal 435bp zone in containing the 50ml reactant mixture of following composition:

-1ml cDNA (producing as mentioned above)

-0.25mM?dNTPs

-0.2mM?PR44(SEQ?ID?NO：58)

-0.2mM?PR45(SEQ?ID?NO：59)

-5ml 10 * PCR buffer solution (TAKARA)

-0.25ml R Taq polymerase (TAKARA)

-28.8ml distilled water.

PCR carries out according to following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

58 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

The pcr amplification reaction that uses primer PR42 and PR43 to carry out produces the 443bp fragment, and uses the pcr amplification reaction of primer PR44 and PR45 to produce the 444bp fragment.

Use standard method that two amplicon PR42-PR43 (HindIII-SalI has a mind to) fragment and PR44-PR45 (EcoRI-BamHI antisense) fragment cloning are gone into PCR cloning vector pCR-BluntII (Invitrogen).Use primer SP6 to carry out sequencing and confirm that its sequence is identical with disclosed sequence A F251016 (SEQ ID NO:38) under each situation, except the restriction enzyme site difference that is imported.Therefore this clone is used for producing oppositely repetition construct (seeing example I .10) at cloning vector pJAI1.

Carry out cloning the first time step by separating 444bp PR44-PR45BamHI-EcoRI fragment from cloning vector pCR-BluntII (Invitrogen) and connecting into the carrier pJAI1 that cuts through the BamHI-EcoRI-enzyme.The clone who contains antisense orientation ε-cyclase 5 '-stub area is called pJAI2.Connect the fusion of transcribing that produces between ε-cyclase 5 '-stub area antisense fragment and the CaMV polyadenylation signal.

Carry out cloning the second time step by separating 443bp PR42-PR43HindIII-SalI fragment from cloning vector pCR-BluntII (Invitrogen) and connecting into the carrier pJAI2 that cuts through the HindIII-SalI-enzyme.Containing intentionally, the clone of direction 435bp ε-cyclase cDNA 5 '-stub area is called pJAI3.Connect to produce AP3P and ε-cyclase 5 '-stub area and have a mind to the fusion of transcribing between the fragment.

In order to be created in the reverse repetition expression cassette under the control of CHRC promoter, utilize petunia (petunia) genomic DNA (producing) and primer PRCHRC5 (SEQID NO:76) and PRCHRC3 (SEQ ID NO:77) amplification CHRC promoter fragment according to standard method.Amplicons cloned is gone into cloning vector pCR2.1 (Invitrogen).Use primer M13 the DCRP pCR2.1-CHRC of institute to be carried out sequencing and confirm that this sequence is identical with sequence A F099501 with T7.Therefore this clone is used to be cloned into expression vector pJAI3.

Clone by separating 1537bp SacI-HindIII fragment from pCR2.1-CHRC and connecting into the carrier pJAI3 that cuts through the SacI-HindIII-enzyme.Contain promoter CHRC but not the clone of original promoter AP3P is called pJCI3.

Utilize binary vector pSUN5 (WO02/00900) generation to be used for carrying out the expression vector of the conversion of the agriculture bacillus mediated reverse repetition transcript under AP3P-or CHRC-control marigold.

In order to produce expression vector pS5AI3, will connect into the carrier pSUN5 (Fig. 8, construct figure) that cuts through the SacI-XhoI-enzyme from the 2622bp SacI-XhoI fragment of pJAI3.

In Fig. 8, Segment A P3P contains modified AP3P promoter (771bp), fragment 5sense contains marigold ε-cyclase 5 ' zone (435bp) of direction intentionally, fragment intron contains potato ST-LS1 gene PIV2 introne, fragment 5anti contains marigold ε-cyclase 5 ' zone (435bp) of antisense orientation, and fragment term (761bp) contains the CaMV polyadenylation signal.

In order to produce expression vector pS5CI3, will connect into the carrier pSUN5 (Fig. 9, construct figure) that cuts through the SacI-XhoI-enzyme from the 3394bp SacI-XhoI fragment of pJCI3.

In Fig. 9, fragment CHRC contains promoter (1537bp), fragment 5sense contains marigold ε-cyclase 5 ' zone (435bp) of direction intentionally, fragment intron contains potato ST-LS1 gene PIV2 introne, fragment 5anti contains marigold ε-cyclase 5 ' zone (435bp) of antisense orientation, and fragment term (761bp) contains the CaMV polyadenylation signal.

Example I .12

Generation is used for the reverse repetition expression cassette (at the 3 ' zone of ε-cyclase cDNA) at marigold specifically expressing ε-cyclase dsRNA

By PCR (PCR) method, use one to have a mind to special primer (PR46SEQ ID NO:60) and an antisense special primer (PR47 SEQ ID NO:61) and increase from marigold and contain ε-cyclase cDNA (Genbank accession number: AF251016) 3 '-stub area (384bp) nucleic acid.Marigold ε-cyclase cDNA3 '-stub area (384bp) is made up of corresponding to the coding region of C end 140bp3 '-non-translated sequence (3 ' UTR) and 244bp.

I.11 prepare total RNA from marigold flower as embodiment as described in.

As embodiment I.11 as described in, use the synthetic cDNA of antisense special primer PR17 (SEQ ID NO:53).

PCR reaction condition subsequently is as follows:

Pcr amplification contains the PR46-PR47 dna fragmentation in ε-cyclase 3 '-terminal 384bp zone in containing the 50ml reactant mixture of following composition:

-1ml cDNA (producing as mentioned above)

-0.25mM?dNTPs

-0.2mM?PR46(SEQ?ID?NO：60)

-0.2mM?PR47(SEQ?ID?NO：61)

-5ml 10 * PCR buffer solution (TAKARA)

-0.25ml R Taq polymerase (TAKARA)

-28.8ml distilled water.

Pcr amplification contains the PR48-PR49 dna fragmentation in ε-cyclase 3 '-terminal 384bp zone in containing the 50ml reactant mixture of following composition:

-1ml cDNA (producing as mentioned above)

-0.25mM?dNTPs

-0.2mM?PR48(SEQ?ID?NO：62)

-0.2mM?PR49(SEQ?ID?NO：63)

-5ml 10 * PCR buffer solution (TAKARA)

-0.25ml R Taq polymerase (TAKARA)

-28.8ml distilled water.

The PCR reaction is carried out according to following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

58 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

Use the fragment of the pcr amplification reaction generation 392bp of SEQ ID NO:60 and SEQ ID NO:61, use the fragment of the pcr amplification reaction generation 396bp of SEQ ID NO:62 and SEQ ID NO:63.

Use standard method with two amplicons, PR46-PR47 fragment and PR48-PR49 fragment are cloned into PCR cloning vector pCR-BluntII (Invitrogen).Use primer SP6 to carry out sequencing and confirm that its sequence is identical with disclosed sequence A F251016 (SEQ ID NO:38) under each situation, except the restriction enzyme site difference that is imported.Therefore described clone is used for producing the reverse repetition construct (seeing example I .10) of cloning vector pJAI1.

Carry out first clone's step by separating 396bp PR48-PR49BamHI-EcoRI fragment from cloning vector pCR-BluntII (Invitrogen) and connecting into the carrier pJAI1 that cuts through the BamHI-EcoRI-enzyme.The clone who contains ε-cyclase 3 '-stub area of antisense orientation is called pJAI4.Connect the fusion of transcribing between the antisense fragment produced ε-cyclase 3 '-stub area and the CaMV polyadenylation signal.

Carry out second clone's step by separating 392bp PR46-PR47HindIII-SalI fragment from cloning vector pCR-BluntII (Invitrogen) and connecting into the carrier pJAI4 that cuts through the HindIII-SalI-enzyme.Containing intentionally, the clone of ε-cyclase the 392bp 3 '-stub area of direction is called pJAI5.Connect and to have produced AP3P and ε-cyclase 3 '-stub area and have a mind to the fusion of transcribing between the fragment.

Use binary vector pSUN5 (WO02/00900) to produce and be used for carrying out the expression vector that the agriculture bacillus mediated reverse repetition transcript under AP3P-control transforms marigold.In order to produce expression vector pS5AI5, will connect into the carrier pSUN5 (Figure 10, construct figure) that cuts through the SacI-XhoI enzyme from the 2523bp SacI-XhoI fragment of pJAI5.

In Figure 10, Segment A P3P contains the AP3P promoter (771bp) of modification, fragment 3sense contains marigold ε-cyclase 3 ' zone (435bp) of direction intentionally, fragment intron contains potato ST-LS1 gene IV2 introne, fragment 3anti contains marigold ε-cyclase 3 ' zone (435bp) of antisense orientation, and fragment term (761bp) contains the CaMV polyadenylation signal.

Example I .13

The clone of ε-cyclase promoter

Use genomic DNA (separating from marigold Orangenprinz strain) by two 199bp fragment and 312bp fragments of independently cloning tactful inverse PCR (adopting Proc.Natl.Acad.SciUSA 90:10370 such as Long) and TAIL-PCR (1995) Plant such as (J.8:457-463) Liu Y-G. separate epsilon-cyclase promoter by standard method.

In containing the 25 μ l reaction mixtures of EcoRV and RsaI, digest 2 μ g genomic DNAs for the inverse PCR method, be diluted to 300 μ l then and use the 3U ligase to reconnect for 16 ℃ and spend the night.Use primer PR50 (SEQ ID NO:64) and PR51 (SEQ ID NO:65), pass through pcr amplification reaction, produced under each situation to have a mind to direction and contained the fragment of 354bp ε-cyclase cDNA (Genbank accession number AF251016), this fragment has been connected (seeing Figure 11) with 5 '-stub area of ε-cyclase cDNA of 300bp ε-cyclase promoter and 70bp.

The PCR reaction condition is as follows:

Pcr amplification contains the PR50-PR51 dna fragmentation of ε-cyclase 312bp promoter fragment in containing the 50ml reaction mixture of following composition:

-1ml connects mixture (producing as mentioned above)

-0.25mM?dNTPs

-0.2mM?PR50(SEQ?ID?NO：64)

-0.2mM?PR51(SEQ?ID?NO：65)

-5ml 10 * PCR buffer solution (TAKARA)

-0.25ml R Taq polymerase (TAKARA)

-28.8ml distilled water.

The PCR reaction is carried out according to following cycling condition:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

53 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

Use the pcr amplification reaction of primer PR50 and PR51 to produce the 734bp fragment, wherein especially contain ε-cyclase 312bp promoter fragment (Figure 11).

Use standard method that amplicons cloned is gone into PCR cloning vector pCR2.1 (Invitrogen), use primer M13 and T7 to carry out sequencing and obtain sequence SEQ ID NO:45.In different amplification experiments, also produced this sequence, so this sequence has been represented the nucleotide sequence among the employed marigold strain Orangenprinz.

For the TAIL-PCR method, under each situation, use different gene specific primers (nested primer) to carry out three continuous PCR reactions.

In containing the 20m1 reaction mixture of following composition, carry out the TAIL1-PCR reaction:

-1ng genomic DNA (producing as mentioned above)

Every kind of dNTP of-0.2mM

-0.2mM?PR60(SEQ?ID?NO：66)

-0.2mM?AD1(SEQ?ID?NO：69)

-2ml 10 * PCR buffer solution (TAKARA)

-0.5U R Taq polymerase (TAKARA)

-mend to 20ml with distilled water.

-AD1 is first mixture of sequence (a/c/g/t) tcga (g/c) t (a/t) t (g/c) g (a/t) gtt primer.

PCR reaction TAIL1 carries out under following cycling condition:

1 * 93 ℃: 1 minute, 95 ℃: 1 minute

5 * 94 ℃: 30 seconds, 62 ℃: 1 minute, 72 ℃: 2.5 minutes

1 * 94 ℃: 30 seconds, 25 ℃: 3 minutes, in 3 minutes, rise to 72 ℃,

72 ℃: 2.5 minutes

15 * 94 ℃: 10 seconds, 68 ℃: 1 minute, 72 ℃: 2.5 minutes;

94 ℃: 10 seconds, 68 ℃: 1 minute, 72 ℃: 2.5 minutes;

94 ℃: 10 seconds, 29 ℃: 1 minute, 72 ℃: 2.5 minutes

1 * 72 ℃: 5 minutes

In containing the 21ml reactant mixture of following composition, carry out TAIL2-PCR:

1: 50 dilution of-1ml TAIL1 reactant mixture (producing as mentioned above)

-0.8mM?dNTP

-0.2mM?PR61(SEQ?ID?NO：67)

-0.2mM?AD1(SEQ?ID?NO：69)

-2ml 10 * PCR buffer solution (TAKARA)

-0.5U R Taq polymerase (TAKARA)

-mend to 21ml with distilled water.

PCR reaction TAIL2 carries out under following cycling condition:

12 * 94 ℃: 10 seconds, 64 ℃: 1 minute, 72 ℃: 2.5 minutes;

94 ℃: 10 seconds, 64 ℃: 1 minute, 72 ℃: 2.5 minutes;

94 ℃: 10 seconds, 29 ℃: 1 minute, 72 ℃: 2.5 minutes

1 * 72 ℃: 5 minutes

In containing the 100ml reactant mixture of following composition, carry out the TAIL3-PCR reaction:

1: 10 dilution of-1ml TAIL2 reactant mixture (producing as mentioned above)

-0.8mM?dNTP

-0.2mM?PR63(SEQ?ID?NO：68)

-0.2mM?AD1(SEQ?ID?NO：69)

-10ml 10 * PCR buffer solution (TAKARA)

-0.5U R Taq polymerase (TAKARA)

-mend to 100ml with distilled water.

PCR reaction TAIL3 carries out under following cycling condition:

20 * 94 ℃: 15 seconds, 29 ℃: 30 seconds, 72 ℃: 2 minutes

1 * 72 ℃: 5 minutes

Use the pcr amplification reaction of primer PR63 and AD1 to produce the 280bp fragment, wherein especially contain ε-cyclase 199bp promoter fragment (Figure 12).

Use standard method that amplicons cloned is gone into PCR cloning vector pCR2.1 (Invitrogen).Use primer M13 and T7 to carry out sequencing and obtain sequence SEQ ID NO:46.This sequence is identical with the sequence SEQ ID NO:45 that separates by the IPCR strategy, has therefore represented the nucleotide sequence among the employed marigold strain Orangenprinz.

The pCR2.1 clone who contains the ε-cyclase promoter 312bp fragment (SEQ IDNO:45) of separating by the IPCR strategy is called pTA-ecycP and is used to produce the IR construct.

Example I .14

Generation is used for the reverse repetition expression cassette (at ε-cyclase cDNA promoter region) at marigold specifically expressing ε-cyclase dsRNA

Under the control of the modified forms AP3P that is expressed in arabidopsis cauliflower specific promoter AP3 of reverse repetition transcript in marigold that forms by ε-cyclase promoter fragment (seeing example I .10) or under the control of flower specific promoter CHRC (Genbank accession number AF099501), carry out.Under each situation, oppositely repeat a ε-cyclase promoter fragment (fragment intentionally) and a reverse ε-cyclase promoter fragment (antisense fragment) (seeing example I .10) that sequence is identical that transcript contains a correct direction that links together by the function introne.

Use DNA (clone pTA-ecycP sees example I .13) and primer PR124 (SEQ ID NO:70) to produce promoter fragment with PR127 (SEQ ID NO:73) by PCR method with PR126 (SEQ ID NO:72) and primer PR125 (SEQ ID NO:71).

The PCR reaction condition is as follows:

Pcr amplification contains the PR124-PR126 dna fragmentation of ε-cyclase promoter fragment in containing the 50ml reactant mixture of following composition:

-1ml cDNA (producing as mentioned above)

-0.25mM?dNTPs

-0.2mM?PR124(SEQ?ID?NO：70)

-0.2mM?PR126(SEQ?ID?NO：72)

-5ml 10 * PCR buffer solution (TAKARA)

-0.25ml R Taq polymerase (TAKARA)

-28.8ml distilled water.

Pcr amplification contains the PR125-PR127 dna fragmentation of ε-cyclase 312bp promoter fragment in containing the 50ml reactant mixture of following composition:

-1ml cDNA (producing as mentioned above)

-0.25mM?dNTPs

-0.2mM?PR125(SEQ?ID?NO：71)

-0.2mM?PR127(SEQ?ID?NO：73)

-5ml 10 * PCR buffer solution (TAKARA)

-0.25ml R Taq polymerase (TAKARA)

-28.8ml distilled water.

Under following cycling condition, carry out the PCR reaction:

1 * 94 ℃ 2 minutes

35 * 94 ℃ 1 minute

53 ℃ 1 minute

72 ℃ 1 minute

1 * 72 ℃ 10 minutes

Use the pcr amplification reaction of primer PR124 and PR126 to produce the 358bp fragment, use the pcr amplification reaction of primer PR125 and PR127 to produce the 361bp fragment.

Use standard method with two amplicons, PR124-PR126 (HindIII-SalI has a mind to) fragment and PR125-PR127 (EcoRI-BamHI antisense) fragment are cloned into PCR cloning vector pCR-BluntII (Invitrogen).Use primer SP6 to carry out sequencing and confirm that its sequence is identical with SEQ ID NO:45 except the restriction enzyme site that is imported under each situation.Therefore this clone is used for producing the reverse repetition construct (seeing example I .10) of cloning vector pJAI1.

Carry out cloning the first time step by separating 358bp PR124-PR126HindIII-SalI fragment from cloning vector pCR-BluntII (Invitrogen) and connecting into the carrier pJAI1 that cuts through the BamHI-EcoRI enzyme.Containing intentionally, the clone of direction ε-cyclase promoter fragment is called cs43.Coupled reaction imports ε-cyclase promoter and has a mind to fragment between AP3P promoter and introne.

Carry out cloning the second time step by separating 361bp PR125-PR127BamHI-EcoRI fragment from cloning vector pCR-BluntII (Invitrogen) and connecting into the carrier cs43 that cuts through the BamHI-EcoRI-enzyme.The clone who contains antisense orientation ε-cyclase promoter fragment is called cs44.Coupled reaction forms between introne and ε-cyclase promoter antisense fragment and transcribes fusion.

In order to be created in the reverse repetition expression cassette under the control of CHRC promoter, use petunia genomic DNA (producing) and primer PRCHRC3 ' (SEQ ID NO:77) and PRCHRC5 ' (SEQ ID NO:76) amplification CHRC promoter fragment according to standard method.Amplicons cloned is gone into cloning vector pCR2.1 (Invitrogen).Use primer M13 the DCRP pCR2.1-CHRC of institute to be carried out sequencing and confirm that this sequence is identical with sequence A F099501 with T7.Therefore this clone is used to be cloned into expression vector cs44.

Clone by separating 1537bp SacI-HindIII fragment from pCR2.1-CHRC and connecting into the carrier cs44 that cuts through the SacI-HindIII enzyme.Contain promoter CHRC but not the clone of original promoter AP3P is called cs45.

In order to be created in the reverse repetition expression cassette under the control of two promoter CHRC promoters and AP3P promoter, the AP3P promoter is cloned into 3 ' end of ε-cyclase antisense fragment of cs45 with antisense orientation.Use the AP3P promoter fragment of primer PR128 and PR129 amplification from pJAI1.Amplicons cloned is gone into cloning vector pCR2.1 (Invitrogen).Use primer M13 to carry out sequencing and confirm that this sequence is identical with sequence SEQ ID NO:28 (AL132971) with T7.This clone pCR2.1-AP3PSX is used to be created in two reverse repetition expression cassettes under the promoter control.

Clone by separating 771bp SalI-XhoI fragment from pCR2.1-AP3PSX and connecting into the carrier cs45 that cuts through the XhoI-enzyme.The clone who contains antisense orientation promoter AP3P in inverted repeats 3 ' side is called cs46.

Use binary vector pSUN5 (WO 02/00900) to produce and be used for carrying out the expression vector that the agriculture bacillus mediated reverse repetition transcript under AP3P-control transforms marigold.

In order to produce expression vector pS5AI7, will connect into the carrier pSUN5 (Figure 13, construct figure) that cuts through the SacI-XhoI-enzyme from the 1685bp SacI-XhoI fragment of cs44.In Figure 13, Segment A P3P contains the AP3P promoter (771bp) of modification, fragment P-sense contains the ε-cyclase 312bp promoter fragment of direction intentionally, fragment intron contains potato ST-LS1 gene IV2 introne, and fragment P-anti contains the ε-cyclase 312bp promoter fragment of antisense orientation.

In order to produce expression vector pS5CI7, will connect into the carrier pSUN5 (Figure 14, construct figure) that cuts through the SacI-XhoI-enzyme from the 2445bp SacI-XhoI fragment of cs45.

In Figure 14, fragment CHRC contains CHRC promoter (1537bp), fragment P-sense contains the ε-cyclase 312bp promoter fragment of direction intentionally, fragment intron contains potato ST-LS1 gene IV2 introne, and fragment P-anti contains the ε-cyclase 312bp promoter fragment of antisense orientation.

In order to produce expression vector pS5CAI7, will connect into the carrier pSUN5 (Figure 15, construct figure) that cuts through the SacI-XhoI-enzyme from the 3219bp SacI-XhoI fragment of cs46.

In Figure 15, fragment CHRC contains CHRC promoter (1537bp), fragment P-sense contains the ε-cyclase 312bp promoter fragment of direction intentionally, fragment intron contains potato ST-LS1 gene IV2 introne, fragment P-anti contains the ε-cyclase 312bp promoter fragment of antisense orientation, and Segment A P3P contains antisense orientation 771bp AP3P promoter fragment.

Example I .15

Generation with transgenosis marigold plant of the ε-cyclase activity that has reduced

Be positioned over (MS culture medium on the germination culture medium with the sterilization of marigold seed and with it; Murashige and Skoog, Physiol.Plant.15 (1962), 473-497) pH 5.8,2% sucrose).It is under the 18-28 ℃/20-200mE/3-16 week condition that germination betides temperature/illumination/time interval, is preferably 21 ℃, 20-70mE, 4-8 week.

Till that time results in all leaves of ectogenetic plant and transverse cuts to middle arteries and veins.In preparation process, be 10-60mm with resulting size ²Leaf explant to be stored in the liquid MS medium room temperature the longest 2 hours.

Use binary vector pS5AI3 to transform Agrobacterium tumefaciems bacterial strain EHA105.The Agrobacterium tumefaciems bacterial strain EHA105 that transforms is grow overnight under the following conditions: monospecific polyclonal is inoculated in YEB (0.1% yeast extract, 0.5% beef extract, 0.5% peptone, 0.5% sucrose, the 0.5% magnesium sulfate 7H that contains the 25mg/l kanamycins ₂O) cultivated 16-20 hour in and at 28 ℃.Pass through then 6000g centrifugal 10 minutes results bacterial suspension and with OD ₆₀₀Approximately the concentration of 0.1-0.8 is resuspended in the liquid MS medium.This suspension is used for cultivating altogether with the leaf material.

Before cultivating altogether, replace preserving the MS culture medium of leaf immediately with bacterial suspension.Gently vibration down room temperature leaf was hatched in transforming the Agrobacterium suspension 30 minutes.Then the explant that infects being positioned over agar solidifies and (contains growth regulator for example on the MS culture medium of 3mg/l benayl aminopurine (BAP) and 1mg/l heteroauxin (IAA) as 0.8% plant agar (Duchefa, NL)).The location of leaf on culture medium is not important.Explant is cultivated 1-8 days, but be preferably 6 days, can use following condition: luminous intensity: 30-80mmol/m ²X second, temperature: 22-24 ℃, light/secretly be replaced by 16/8 hour.Then, the explant of cultivating altogether is transferred to fresh MS culture medium, is preferably the fresh MS culture medium that contains the isometric growth conditioning agent, this second culture medium additionally contains the antibiotic of bacteria growing inhibiting.Concentration is that the Ticarcillin/Clavulanate Acid of 200-500mg/l is particularly suitable for this purpose.As second selection component, spendable is to be used to screen the composition that successfully transforms.Concentration is that the phosphinothricin selection of 1-5mg/l is very effective, but also can consider to use other selection component according to this method.

After cultivating for one to three week under each situation, explant is transferred in the fresh culture up to plumule and budlet forms, then they are transferred to and contain the composition that Ticarcillin/Clavulanate Acid and PPT or other contain conditioning agent and just refer to such as 0.5mg/l indolebutyric acid (IBA) and 0.5mg/l gibberellic acid GA ₃The same basic culture medium in to take root.The bud of taking root can be transferred to the greenhouse.

Except said method, following promising change also is possible:

Before the bacterial infection explant, for cultivation altogether can be preferably 3-4 days with explant in above-mentioned culture medium preincubate 1-12 days.Then, infect as mentioned above, cultivate altogether and selectivity regeneration.

According to above-mentioned method for transformation, use expression construct pS5AI3, obtain following strain:

CS30-1, CS30-3 and CS30-4

Example I .16:

Characteristic with transgenosis marigold plant of the ε-cyclase activity that has reduced

In liquid nitrogen, grind transgenosis marigold plant floral material with mortar from example I .15, and with 100% acetone extracting (three times, each 500ml) powder (approximately 250-500mg).Evaporating solvent also is resuspended in 100ml acetone with carotenoid.

Use C30 reversed-phase column method, quantitatively different carotenoid.In fact (Frazer etc. (2000), Plant Journal 24 (4): 551-558) identical with disclosed method for the HPLC condition of work.Can identify carotenoid based on UV-VIS spectrum.

Table 2 has shown carotenoid spectrum in transgenosis marigold plant that produces according to the foregoing description and the marigold petal that contrasts marigold plant.The amount of all carotenoid provides with [μ g/g] fresh weight, changes at the parenthetic percentage of comparing with check plant that provides.

Compare with the check plant of non-genetic modification, has the carotenoid content that the genetically modified plant of the ε-cyclase activity that has reduced has " the beta carotene approach " of remarkable increase, the beta carotene and the luteole content that have for example significantly increased, and the carotenoid content of " the alpha-carotene approach " that significantly reduced, the lutein content that has for example significantly reduced.

Table 2

Plant	Lutein	Beta carotene	Luteole	Violaxanthin	Total carotinoid
Plant	Lutein	Beta carotene	Luteole	Violaxanthin	Total carotinoid	Contrast	??260	4.8	2.7	36	?304
??CS?30-1	??35(-86％)	13(+170％)	4.4(+62％)	59(+63％)	?111(-63％)	Contrast	??260	4.8	2.7	36	?304
??CS?30-1	??35(-86％)	13(+170％)	4.4(+62％)	59(+63％)	?111(-63％)	Contrast	??456	6.4	6.9	58	?527
??CS?30-3	??62(-86％)	13(+103％)	8.9(+29％)	75(+29％)	?159(-70％)	Contrast	??456	6.4	6.9	58	?527
??CS?30-3	??62(-86％)	13(+103％)	8.9(+29％)	75(+29％)	?159(-70％)	??CS?30-4	??68(-85％)	9.1(+42％)	5.7(-17％)	61(+5％)	?144(-73％)

Example II

The generation of the plants of tagetes species part of astaxanthin-containing

Isolate the capitulum or the petal and dry of the astaxanthin-containing plants of tagetes species that produces according to example I .6.Capitulum or petal with drying changes into powder type by pulverizing then.

EXAMPLE III

The generation of astaxanthin-containing extract and being further purified

In homogenizer with excessive (the about 10 parts of solvents of a vegetable material) solvent (for example acetone, hexane, carrene, methyl tertiary butyl ether(MTBE), oxolane, ethanol, heptane, cycloheptane or benzinum, but be not limited in these) or with solvent mixture (for example acetone/hexane, ethanol/hexane are (50: 50, v/v) or acetone/methanol (7: 3, v/v)) dry floral leaf or the dry capitulum of the marigold that produces according to example I .6 of homogenate, and vibration is down in dark and the extracting of cold place.Residue can be repeated extracting nearly three times with employed solvent/solvents mixture.Use collected organic solvent of evaporator evaporation or solvent mixture up to the concentrate that has obtained reducing.In addition, can be with the further extracting material of hexane.The employed hexane of evaporation (also at dark and cold place).

To be dissolved in hexane with the concentrate that this kind mode produces and utilize column chromatography to use earth silicon material to carry out chromatography.A copy of it earth silicon material is mixed with 1-2 part carotenoid solutions and be loaded in the post.At dark and cold place with the abundant column scrubber of hexane.Abandon this eluate.With hexane and alcohol mixture (2-5% ethanol is arranged in the hexane) wash-out keto-acid carotenoid astaxanthin particularly, go out orange red fraction up to wash-out.Collect orange red eluate up to color change.Orange red eluate contains monoesters and diester astaxanthin mixture.

EXAMPLE IV

The generation of extrusion pressing type trout feed, described feed comprise plants of tagetes species or plant part that contains astaxanthin or the astaxanthin-containing extract that contains astaxanthin plants of tagetes species or plant part

The following composition of extruding in double-screw extrusion device.

Composition	????(％)	Weight kg in 500kg
Composition	????(％)	Weight kg in 500kg	Wheaten starch premix vitamin Lipotril (50%) the wheat gluten Sipernat 50S fish oil of in advance gel of fish meal full-fat bean	????30.00 ????20.00 ????18.00 ????0.80 ????0.20 ????20.00 ????3.00 ????8.00	????150.00 ????100.00 ????90.00 ????4.00 ????1.00 ????100.00 ????15.00 ????40.00

Before the extruding, as plants of tagetes species or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part of component adding according to the astaxanthin-containing that produces such as example II through pulverizing processing.

After the extruding, with the astaxanthin-containing extract of the plants of tagetes species of astaxanthin-containing or plant part or through the processing extract with liquid form, for example produce according to EXAMPLE III, sparge on the extrudate and (use) by the PPA method.

The dosage of astaxanthin active component is to contain 10,20 and the 40mg astaxanthin in every kg feed.

Finish after the extrusion process dry and cooling with extrudate.

EXAMPLE V

The astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part is administered orally in trout-bioavilability with the standard trout form of feeding to be detected

The trout feed that contains astaxanthin pigment of the present invention that produces according to EXAMPLE IV also is administered orally in trout (average live body quality 180g).Test 3 concentration: every kg feed 10,20 and 40mg are from the astaxanthin of astaxanthin pigment of the present invention.

As culturing trout described in hereinafter:

● give 14 day laundering period of trout standard.

● during feeding experiment, in the permanent current plastics cylinder of each volume 80 premium on currency, culture 10 tail trouts.Water temperature is 15 ℃.Biology purification of water process and every day, the water of cumulative volume 10% replaced with fresh water at least.

● the illumination phase is that 12 hours every days are to avoid the too early sexal maturity of animal.

● each is handled, and the quantity in pond is 3.This equals each dosage level 30 tail trout.

● feed storage in-20 ℃ to avoid astaxanthin loss.Thawing feed a part (weekly) is also used.

● experiment periods was 8 weeks.

Feed and raise as carrying out trout described in hereinafter:

● the experiment feed of being used is the extrusion pressing type trout feed of having carried out oily dressing in addition that produces according to EXAMPLE IV.

● during the laundering period, the extrusion pressing type oil dressing of using the no astaxanthin that produces according to EXAMPLE IV does not have astaxanthin standard trout feed.

● as negative control, the extrusion pressing type oil dressing that whole experimental session is used the no astaxanthin that produces according to EXAMPLE IV does not have astaxanthin standard trout feed.

● every day 2 * carry out feeding with hand to have enough up to animal.

The painted behavior parameter that not only influences fish of astaxanthin of the present invention, for example feed picked-up, feed conversion and body weight increase, but also the painted biologicak efficiency that influence is studied.

Statistical evaluation is carried out in average food consumption, food conversion and body weight increase to every tail fish.

Compare under each situation the content astaxanthin in the fillet (mg/kg) with negative control and measure the painted of fish by reflective spectrophotometer determination method (red value of Minolta a value=fillet incision) and by measuring.

Represent the Minolta a value of the red part in the tone to increase with of the reduction of dosage dependence mode with the function ladder.The Minolta b value of reflection yl moiety is near negative scope or zero.This shows that the red color tone in the trout fillet depends on the amount of the astaxanthin that is consumed.

Experimental session, for observed behavior parameter, (extract of the powder of astaxanthin-containing, liquid form astaxanthin-containing, synthesizing astaxanthin, negative control) do not observed statistics and gone up significant difference between processed group or in the processed group.

The astaxanthin-containing extract of finding the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part is that the trout of representative is biological available and the trout biological behaviour do not caused negative effect in painted making with the salmonid.

Sequence table

＜110〉Sungene GmbH ﹠ Co. KG

＜120〉plants of tagetes species of astaxanthin-containing or plant part are as the purposes of feed

<130>PF?54148

<160>142

＜170〉PatentIn version 3 .1

<210>1

<211>1771

<212>DNA

＜213〉haematococcus pluvialis (Haematococcus pluvialis)

<220>

<221>CDS

<222>(166)..(1155)

<223>

<400>1

ggcacgagct?tgcacgcaag?tcagcgcgcg?caagtcaaca?cctgccggtc?cacagcctca??????60

aataataaag?agctcaagcg?tttgtgcgcc?tcgacgtggc?cagtctgcac?tgccttgaac?????120

ccgcgagtct?cccgccgcac?tgactgccat?agcacagcta?gacga?atg?cag?cta?gca?????177

Met?Gln?Leu?Ala

1

gcg?aca?gta?atg?ttg?gag?cag?ctt?acc?gga?agc?gct?gag?gca?ctc?aag???????225

Ala?Thr?Val?Met?Leu?Glu?Gln?Leu?Thr?Gly?Ser?Ala?Glu?Ala?Leu?Lys

5???????????????????10??????????????????15??????????????????20

gag?aag?gag?aag?gag?gtt?gca?ggc?agc?tct?gac?gtg?ttg?cgt?aca?tgg???????273

Glu?Lys?Glu?Lys?Glu?Val?Ala?Gly?Ser?Ser?Asp?Val?Leu?Arg?Thr?Trp

25??????????????????30??????????????????35

gcg?acc?cag?tac?tcg?ctt?ccg?tca?gaa?gag?tca?gac?gcg?gcc?cgc?ccg???????321

Ala?Thr?Gln?Tyr?Ser?Leu?Pro?Ser?Glu?Glu?Ser?Asp?Ala?Ala?Arg?Pro

40??????????????????45??????????????????50

gga?ctg?aag?aat?gcc?tac?aag?cca?cca?cct?tcc?gac?aca?aag?ggc?atc???????369

Gly?Leu?Lys?Asn?Ala?Tyr?Lys?Pro?Pro?Pro?Ser?Asp?Thr?Lys?Gly?Ile

55???????????????????60??????????????????65

aca?atg?gcg?cta?cgt?gtc?atc?ggc?tcc?tgg?gcc?gca?gtg?ttc?ctc?cac???????417

Thr?Met?Ala?Leu?Arg?Val?Ile?Gly?Ser?Trp?Ala?Ala?Val?Phe?Leu?His

70??????????????????75??????????????????80

gcc?att?ttt?caa?atc?aag?ctt?ccg?acc?tcc?ttg?gac?cag?ctg?cac?tgg???????465

Ala?Ile?Phe?Gln?Ile?Lys?Leu?Pro?Thr?Ser?Leu?Asp?Gln?Leu?His?Trp

85??????????????????90??????????????????95??????????????????100

ctg?ccc?gtg?tca?gat?gcc?aca?gct?cag?ctg?gtt?agc?ggc?acg?agc?agc???????513

Leu?Pro?Val?Ser?Asp?Ala?Thr?Ala?Gln?Leu?Val?Ser?Gly?Thr?Ser?Ser

105?????????????????110?????????????????????115

ctg?ctc?gac?atc?gtc?gta?gta?ttc?ttt?gtc?ctg?gag?ttc?ctg?tac?aca???????561

Leu?Leu?Asp?Ile?Val?Val?Val?Phe?Phe?Val?Leu?Glu?Phe?Leu?Tyr?Thr

120?????????????????125?????????????????130

ggc?ctt?ttt?atc?acc?acg?cat?gat?gct?atg?cat?ggc?acc?atc?gcc?atg???????609

Gly?Leu?Phe?Ile?Thr?Thr?His?Asp?Ala?Met?His?Gly?Thr?Ile?Ala?Met

135?????????????????140?????????????????145

aga?aac?agg?cag?ctt?aat?gac?ttc?ttg?ggc?aga?gta?tgc?atc?tcc?ttg???????657

Arg?Asn?Arg?Gln?Leu?Asn?Asp?Phe?Leu?Gly?Arg?Val?Cys?Ile?Ser?Leu

150??????????????????155?????????????????160

tac?gcc?tgg?ttt?gat?tac?aac?atg?ctg?cac?cgc?aag?cat?tgg?gag?cac???????705

Tyr?Ala?Trp?Phe?Asp?Tyr?Asn?Met?Leu?His?Arg?Lys?His?Trp?Glu?His

165?????????????????170?????????????????175?????????????????180

cac?aac?cac?act?ggc?gag?gtg?ggc?aag?gac?cct?gac?ttc?cac?agg?gga???????753

His?Asn?His?Thr?Gly?Glu?Val?Gly?Lys?Asp?Pro?Asp?Phe?His?Arg?Gly

185?????????????????190?????????????????195

aac?cct?ggc?att?gtg?ccc?tgg?ttt?gcc?agc?ttc?atg?tcc?agc?tac?atg???????801

Asn?Pro?Gly?Ile?Val?Pro?Trp?Phe?Ala?Ser?Phe?Met?Ser?Ser?Tyr?Met

200?????????????????205?????????????????210

tcg?atg?tgg?cag?ttt?gcg?cgc?ctc?gca?tgg?tgg?acg?gtg?gtc?atg?cag???????849

Ser?Met?Trp?Gln?Phe?Ala?Arg?Leu?Ala?Trp?Trp?Thr?Val?Val?Met?Gln

215?????????????????220?????????????????225

ctg?ctg?ggt?gcg?cca?atg?gcg?aac?ctg?ctg?gtg?ttc?atg?gcg?gcc?gcg???????897

Leu?Leu?Gly?Ala?Pro?Met?Ala?Asn?Leu?Leu?Val?Phe?Met?Ala?Ala?Ala

230?????????????????235?????????????????240

ccc?atc?ctg?tcc?gcc?ttc?cgc?ttg?ttc?tac?ttt?ggc?acg?tac?atg?ccc???????945

Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu?Phe?Tyr?Phe?Gly?Thr?Tyr?Met?Pro

245?????????????????250?????????????????255?????????????????260

cac?aag?cct?gag?cct?ggc?gcc?gcg?tca?ggc?tct?tca?cca?gcc?gtc?atg???????993

His?Lys?Pro?Glu?Pro?Gly?Ala?Ala?Ser?Gly?Ser?Ser?Pro?Ala?Val?Met

265?????????????????270?????????????????275

aac?tgg?tgg?aag?tcg?cgc?act?agc?cag?gcg?tcc?gac?ctg?gtc?agc?ttt??????1041

Asn?Trp?Trp?Lys?Ser?Arg?Thr?Ser?Gln?Ala?Ser?Asp?Leu?Val?Ser?Phe

280?????????????????285?????????????????290

ctg?acc?tgc?tac?cac?ttc?gac?ctg?cac?tgg?gag?cac?cac?cgc?tgg?ccc??????1089

Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?His?Trp?Glu?His?His?Arg?Trp?Pro

295?????????????????300?????????????????305

ttc?gcc?ccc?tgg?tgg?gag?ctg?ccc?aac?tgc?cgc?cgc?ctg?tct?ggc?cga??????1137

Phe?Ala?Pro?Trp?Trp?Glu?Leu?Pro?Asn?Cys?Arg?Arg?Leu?Ser?Gly?Arg

310?????????????????315?????????????????320

ggt?ctg?gtt?cct?gcc?tag?ctggacacac?tgcagtgggc?cctgctgcca?????????????1185

Gly?Leu?Val?Pro?Ala

325

gctgggcatg?caggttgtgg?caggactggg?tgaggtgaaa?agctgcaggc?gctgctgccg????1245

gacacgctgc?atgggctacc?ctgtgtagct?gccgccacta?ggggaggggg?tttgtagctg????1305

tcgagcttgc?cccatggatg?aagctgtgta?gtggtgcagg?gagtacaccc?acaggccaac????1365

acccttgcag?gagatgtctt?gcgtcgggag?gagtgttggg?cagtgtagat?gctatgattg????1425

tatcttaatg?ctgaagcctt?taggggagcg?acacttagtg?ctgggcaggc?aacgccctgc????1485

aaggtgcagg?cacaagctag?gctggacgag?gactcggtgg?caggcaggtg?aagaggtgcg????1545

ggagggtggt?gccacaccca?ctgggcaaga?ccatgctgca?atgctggcgg?tgtggcagtg????1605

agagctgcgt?gattaactgg?gctatggatt?gtttgagcag?tctcacttat?tctttgatat????1665

agatactggt?caggcaggtc?aggagagtga?gtatgaacaa?gttgagaggt?ggtgcgctgc????1725

ccctgcgctt?atgaagctgt?aacaataaag?tggttcaaaa?aaaaaa???????????????????1771

<210>2

<211>329

<212>PRT

＜213〉haematococcus pluvialis

<400>2

Met?Gln?Leu?Ala?Ala?Thr?Val?Met?Leu?Glu?Gln?Leu?Thr?Gly?Ser?Ala

1???????????????5???????????????????10??????????????????15

Glu?Ala?Leu?Lys?Glu?Lys?Glu?Lys?Glu?Val?Ala?Gly?Ser?Ser?Asp?Val

20??????????????????25??????????????????30

Leu?Arg?Thr?Trp?Ala?Thr?Gln?Tyr?Ser?Leu?Pro?Ser?Glu?Glu?Ser?Asp

35??????????????????40??????????????????45

Ala?Ala?Arg?Pro?Gly?Leu?Lys?Asn?Ala?Tyr?Lys?Pro?Pro?Pro?Ser?Asp

50??????????????????55??????????????????60

Thr?Lys?Gly?Ile?Thr?Met?Ala?Leu?Arg?Val?Ile?Gly?Ser?Trp?Ala?Ala

65??????????????????70??????????????????75??????????????????80

Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile?Lys?Leu?Pro?Thr?Ser?Leu?Asp

85??????????????????90??????????????????95

Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Asp?Ala?Thr?Ala?Gln?Leu?Val?Ser

100?????????????????105?????????????????110

Gly?Thr?Ser?Ser?Leu?Leu?Asp?Ile?Val?Val?Val?Phe?Phe?Val?Leu?Glu

115?????????????????120?????????????????125

Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Thr?His?Asp?Ala?Met?His?Gly

130?????????????????135?????????????????140

Thr?Ile?Ala?Met?Arg?Asn?Arg?Gln?Leu?Asn?Asp?Phe?Leu?Gly?Arg?Val

145?????????????????150?????????????????155?????????????????160

Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp?Tyr?Asn?Met?Leu?His?Arg?Lys

165?????????????????170?????????????????175

His?Trp?Glu?His?His?Asn?His?Thr?Gly?Glu?Val?Gly?Lys?Asp?Pro?Asp

180?????????????????185?????????????????190

Phe?His?Arg?Gly?Asn?Pro?Gly?Ile?Val?Pro?Trp?Phe?Ala?Ser?Phe?Met

195?????????????????200?????????????????205

Ser?Ser?Tyr?Met?Ser?Met?Trp?Gln?Phe?Ala?Arg?Leu?Ala?Trp?Trp?Thr

210?????????????????215?????????????????220

Val?Val?Met?Gln?Leu?Leu?Gly?Ala?Pro?Met?Ala?Asn?Leu?Leu?Val?Phe

225?????????????????230?????????????????235?????????????????240

Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu?Phe?Tyr?Phe?Gly

245?????????????????250?????????????????255

Thr?Tyr?Met?Pro?His?Lys?Pro?Glu?Pro?Gly?Ala?Ala?Ser?Gly?Ser?Ser

260?????????????????265?????????????????270

Pro?Ala?Val?Met?Asn?Trp?Trp?Lys?Ser?Arg?Thr?Ser?Gln?Ala?Ser?Asp

275?????????????????280?????????????????285

Leu?Val?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?His?Trp?Glu?His

290?????????????????295?????????????????300

His?Arg?Trp?Pro?Phe?Ala?Pro?Trp?Trp?Glu?Leu?Pro?Asn?Cys?Arg?Arg

305?????????????????310?????????????????315?????????????????320

Leu?Ser?Gly?Arg?Gly?Leu?Val?Pro?Ala

325

<210>3

<211>1662

<212>DNA

＜213〉haematococcus pluvialis

<220>

<221>CDS

<222>(168)..(1130)

<223>

<400>3

cggggcaact?caagaaattc?aacagctgca?agcgcgcccc?agcctcacag?cgccaagtga??????60

gctatcgacg?tggttgtgag?cgctcgacgt?ggtccactga?cgggcctgtg?agcctctgcg?????120

ctccgtcctc?tgccaaatct?cgcgtcgggg?cctgcctaag?tcgaaga?atg?cac?gtc???????176

Met?His?Val

1

gca?tcg?gca?cta?atg?gtc?gag?cag?aaa?ggc?agt?gag?gca?gct?gct?tcc???????224

Ala?Ser?Ala?Leu?Met?Val?Glu?Gln?Lys?Gly?Ser?Glu?Ala?Ala?Ala?Ser

5???????????????????10??????????????????15

agc?cca?gac?gtc?ttg?aga?gcg?tgg?gcg?aca?cag?tat?cac?atg?cca?tcc???????272

Ser?Pro?Asp?Val?Leu?Arg?Ala?Trp?Ala?Thr?Gln?Tyr?His?Met?Pro?Ser

20??????????????????25??????????????????30??????????????????35

gag?tcg?tca?gac?gca?gct?cgt?cct?gcg?cta?aag?cac?gcc?tac?aaa?cct???????320

Glu?Ser?Ser?Asp?Ala?Ala?Arg?Pro?Ala?Leu?Lys?His?Ala?Tyr?Lys?Pro

40??????????????????45??????????????????50

cca?gca?tct?gac?gcc?aag?ggc?atc?acg?atg?gcg?ctg?acc?atc?att?ggc???????368

Pro?Ala?Ser?Asp?Ala?Lys?Gly?Ile?Thr?Met?Ala?Leu?Thr?Ile?Ile?Gly

55??????????????????60??????????????????????65

acc?tgg?acc?gca?gtg?ttt?tta?cac?gca?ata?ttt?caa?atc?agg?cta?ccg???????416

Thr?Trp?Thr?Ala?Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile?Arg?Leu?Pro

70??????????????????75??????????????????80

aca?tcc?atg?gac?cag?ctt?cac?tgg?ttg?cct?gtg?tcc?gaa?gcc?aca?gcc???????464

Thr?Ser?Met?Asp?Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Glu?Ala?Thr?Ala

85??????????????????90??????????????????95

cag?ctt?ttg?ggc?gga?agc?agc?agc?cta?ctg?cac?atc?gct?gca?gtc?ttc???????512

Gln?Leu?Leu?Gly?Gly?Ser?Ser?Ser?Leu?Leu?His?Ile?Ala?Ala?Val?Phe

100?????????????????105?????????????????110?????????????????115

att?gta?ctt?gag?ttc?ctg?tac?act?ggt?cta?ttc?atc?acc?aca?cat?gac???????560

Ile?Val?Leu?Glu?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Thr?His?Asp

120?????????????????125?????????????????130

gca?atg?cat?ggc?acc?ata?gct?ttg?agg?cac?agg?cag?ctc?aat?gat?ctc???????608

Ala?Met?His?Gly?Thr?Ile?Ala?Leu?Arg?His?Arg?Gln?Leu?Asn?Asp?Leu

135?????????????????140?????????????????145

ctt?ggc?aac?atc?tgc?ata?tca?ctg?tac?gcc?tgg?ttt?gac?tac?agc?atg???????656

Leu?Gly?Asn?Ile?Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp?Tyr?Ser?Met

150?????????????????155?????????????????160

ctg?cat?cgc?aag?cac?tgg?gag?cac?cac?aac?cat?act?ggc?gaa?gtg?ggg???????704

Leu?His?Arg?Lys?His?Trp?Glu?His?His?Asn?His?Thr?Gly?Glu?Val?Gly

165?????????????????170?????????????????175

aaa?gac?cct?gac?ttc?cac?aag?gga?aat?ccc?ggc?ctt?gtc?ccc?tgg?ttc???????752

Lys?Asp?Pro?Asp?Phe?His?Lys?Gly?Asn?Pro?Gly?Leu?Val?Pro?Trp?Phe

180?????????????????185?????????????????190?????????????????195

gcc?agc?ttc?atg?tcc?agc?tac?atg?tcc?ctg?tgg?cag?ttt?gcc?cgg?ctg???????800

Ala?Ser?Phe?Met?Ser?Ser?Tyr?Met?Ser?Leu?Trp?Gln?Phe?Ala?Arg?Leu

200?????????????????205?????????????????210

gca?tgg?tgg?gca?gtg?gtg?atg?caa?atg?ctg?ggg?gcg?ccc?atg?gca?aat???????848

Ala?Trp?Trp?Ala?Val?Val?Met?Gln?Met?Leu?Gly?Ala?Pro?Met?Ala?Asn

215?????????????????220?????????????????225

ctc?cta?gtc?ttc?atg?gct?gca?gcc?cca?atc?ttg?tca?gca?ttc?cgc?ctc???????896

Leu?Leu?Val?Phe?Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu

230?????????????????235?????????????????240

ttc?tac?ttc?ggc?act?tac?ctg?cca?cac?aag?cct?gag?cca?ggc?cct?gca???????944

Phe?Tyr?Phe?Gly?Thr?Tyr?Leu?Pro?His?Lys?Pro?Glu?Pro?Gly?Pro?Ala

245?????????????????250?????????????????255

gca?ggc?tct?cag?gtg?atg?gcc?tgg?ttc?agg?gcc?aag?aca?agt?gag?gca???????992

Ala?Gly?Ser?Gln?Val?Met?Ala?Trp?Phe?Arg?Ala?Lys?Thr?Ser?Glu?Ala

260?????????????????265?????????????????270?????????????????275

tct?gat?gtg?atg?agt?ttc?ctg?aca?tgc?tac?cac?ttt?gac?ctg?cac?tgg??????1040

Ser?Asp?Val?Met?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?His?Trp

280?????????????????285?????????????????290

gag?cac?cac?agg?tgg?ccc?ttt?gcc?ccc?tgg?tgg?cag?ctg?ccc?cac?tgc??????1088

Glu?His?His?Arg?Trp?Pro?Phe?Ala?Pro?Trp?Trp?Gln?Leu?Pro?His?Cys

295?????????????????300?????????????????305

cgc?cgc?ctg?tcc?ggg?cgt?ggc?ctg?gtg?cct?gcc?ttg?gca?tga??????????????1130

Arg?Arg?Leu?Ser?Gly?Arg?Gly?Leu?Val?Pro?Ala?Leu?Ala

310?????????????????315?????????????????320

cctggtccct?ccgctggtga?cccagcgtct?gcacaagagt?gtcatgctac?agggtgctgc????1190

ggccagtggc?agcgcagtgc?actctcagcc?tgtatggggc?taccgctgtg?ccactgagca????1250

ctgggcatgc?cactgagcac?tgggcgtgct?actgagcaat?gggcgtgcta?ctgagcaatg????1310

ggcgtgctac?tgacaatggg?cgtgctactg?gggtctggca?gtggctagga?tggagtttga????1370

tgcattcagt?agcggtggcc?aacgtcatgt?ggatggtgga?agtgctgagg?ggtttaggca????1430

gccggcattt?gagagggcta?agttataaat?cgcatgctgc?tcatgcgcac?atatctgcac????1490

acagccaggg?aaatcccttc?gagagtgatt?atgggacact?tgtattggtt?tcgtgctatt????1550

gttttattca?gcagcagtac?ttagtgaggg?tgagagcagg?gtggtgagag?tggagtgagt????1610

gagtatgaac?ctggtcagcg?aggtgaacag?cctgtaatga?atgactctgt?ct????????????1662

<210>4

<211>320

<212>PRT

＜213〉haematococcus pluvialis

<400>4

Met?His?Val?Ala?Ser?Ala?Leu?Met?Val?Glu?Gln?Lys?Gly?Ser?Glu?Ala

1???????????????5???????????????????10??????????????????15

Ala?Ala?Ser?Ser?Pro?Asp?Val?Leu?Arg?Ala?Trp?Ala?Thr?Gln?Tyr?His

20??????????????????25??????????????????30

Met?Pro?Ser?Glu?Ser?Ser?Asp?Ala?Ala?Arg?Pro?Ala?Leu?Lys?His?Ala

35??????????????????40??????????????????45

Tyr?Lys?Pro?Pro?Ala?Ser?Asp?Ala?Lys?Gly?Ile?Thr?Met?Ala?Leu?Thr

50??????????????????55??????????????????60

Ile?Ile?Gly?Thr?Trp?Thr?Ala?Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile

65??????????????????70??????????????????75??????????????????80

Arg?Leu?Pro?Thr?Ser?Met?Asp?Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Glu

85??????????????????90??????????????????95

Ala?Thr?Ala?Gln?Leu?Leu?Gly?Gly?Ser?Ser?Ser?Leu?Leu?His?Ile?Ala

100?????????????????105?????????????????110

Ala?Val?Phe?Ile?Val?Leu?Glu?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr

115?????????????????120?????????????????125

Thr?His?Asp?Ala?Met?His?Gly?Thr?Ile?Ala?Leu?Arg?His?Arg?Gln?Leu

130?????????????????135?????????????????140

Asn?Asp?Leu?Leu?Gly?Asn?Ile?Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp

145?????????????????150?????????????????155?????????????????160

Tyr?Ser?Met?Leu?His?Arg?Lys?His?Trp?Glu?His?His?Asn?His?Thr?Gly

165?????????????????170?????????????????175

Glu?Val?Gly?Lys?Asp?Pro?Asp?Phe?His?Lys?Gly?Asn?Pro?Gly?Leu?Val

180?????????????????185?????????????????190

Pro?Trp?Phe?Ala?Ser?Phe?Met?Ser?Ser?Tyr?Met?Ser?Leu?Trp?Gln?Phe

195?????????????????200?????????????????205

Ala?Arg?Leu?Ala?Trp?Trp?Ala?Val?Val?Met?Gln?Met?Leu?Gly?Ala?Pro

210?????????????????215?????????????????220

Met?Ala?Asn?Leu?Leu?Val?Phe?Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala

225?????????????????230?????????????????235?????????????????240

Phe?Arg?Leu?Phe?Tyr?Phe?Gly?Thr?Tyr?Leu?Pro?His?Lys?Pro?Glu?Pro

245?????????????????250?????????????????255

Gly?Pro?Ala?Ala?Gly?Ser?Gln?Val?Met?Ala?Trp?Phe?Arg?Ala?Lys?Thr

260?????????????????265??????????????????270

Ser?Glu?Ala?Ser?Asp?Val?Met?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp

275?????????????????280?????????????????285

Leu?His?Trp?Glu?His?His?Arg?Trp?Pro?Phe?Ala?Pro?Trp?Trp?Gln?Leu

290?????????????????295?????????????????300

Pro?His?Cys?Arg?Arg?Leu?Ser?Gly?Arg?Gly?Leu?Val?Pro?Ala?Leu?Ala

305?????????????????310?????????????????315?????????????????320

<210>5

<211>729

<212>DNA

＜213〉orange Agrobacterium (Agrobacterium aurantiacum)

<220>

<221>CDS

<222>(1)..(729)

<223>

<400>5

atg?agc?gca?cat?gcc?ctg?ccc?aag?gca?gat?ctg?acc?gcc?acc?agc?ctg????????48

Met?Ser?Ala?His?Ala?Leu?Pro?Lys?Ala?Asp?Leu?Thr?Ala?Thr?Ser?Leu

1???????????????5???????????????????10??????????????????15

atc?gtc?tcg?ggc?ggc?atc?atc?gcc?gct?tgg?ctg?gcc?ctg?cat?gtg?cat????????96

Ile?Val?Ser?Gly?Gly?Ile?Ile?Ala?Ala?Trp?Leu?Ala?Leu?His?Val?His

20??????????????????25??????????????????30

gcg?ctg?tgg?ttt?ctg?gac?gca?gcg?gcg?cat?ccc?atc?ctg?gcg?atc?gca???????144

Ala?Leu?Trp?Phe?Leu?Asp?Ala?Ala?Ala?His?Pro?Ile?Leu?Ala?Ile?Ala

35??????????????????40??????????????????45

aat?ttc?ctg?ggg?ctg?acc?tgg?ctg?tcg?gtc?gga?ttg?ttc?atc?atc?gcg???????192

Asn?Phe?Leu?Gly?Leu?Thr?Trp?Leu?Ser?Val?Gly?Leu?Phe?Ile?Ile?Ala

50??????????????????55??????????????????60

cat?gac?gcg?atg?cac?ggg?tcg?gtg?gtg?ccg?ggg?cgt?ccg?cgc?gcc?aat???????240

His?Asp?Ala?Met?His?Gly?Ser?Val?Val?Pro?Gly?Arg?Pro?Arg?Ala?Asn

65??????????????????70??????????????????75??????????????????80

gcg?gcg?atg?ggc?cag?crt?gtc?ctg?tgg?ctg?tat?gcc?gga?ttt?tcg?tgg???????288

Ala?Ala?Met?Gly?Gln?Leu?Val?Leu?Trp?Leu?Tyr?Ala?Gly?Phe?Ser?Trp

85??????????????????90??????????????????95

cgc?aag?atg?atc?gtc?aag?cac?atg?gcc?cat?cac?cgc?cat?gcc?gga?acc???????336

Arg?Lys?Met?Ile?Val?Lys?His?Met?Ala?His?His?Arg?His?Ala?Gly?Thr

100?????????????????105?????????????????110

gac?gac?gac?ccc?gat?ttc?gac?cat?ggc?ggc?ccg?gtc?cgc?tgg?tac?gcc???????384

Asp?Asp?Asp?Pro?Asp?Phe?Asp?His?Gly?Gly?Pro?Val?Arg?Trp?Tyr?Ala

115?????????????????120?????????????????125

cgc?ttc?atc?ggc?acc?tat?ttc?ggc?tgg?cgc?gag?ggg?ctg?ctg?ctg?ccc???????432

Arg?Phe?Ile?Gly?Thr?Tyr?Phe?Gly?Trp?Arg?Glu?Gly?Leu?Leu?Leu?Pro

130?????????????????135?????????????????140

gtc?atc?gtg?acg?gtc?tat?gcg?ctg?atc?ctt?ggg?gat?cgc?tgg?atg?tac???????480

Val?Ile?Val?Thr?Val?Tyr?Ala?Leu?Ile?Leu?Gly?Asp?Arg?Trp?Met?Tyr

145?????????????????150?????????????????155?????????????????160

gtg?gtc?ttc?tgg?ccg?ctg?ccg?tcg?atc?ctg?gcg?tcg?atc?cag?ctg?ttc???????528

Val?Val?Phe?Trp?Pro?Leu?Pro?Ser?Ile?Leu?Ala?Ser?Ile?Gln?Leu?Phe

165?????????????????170?????????????????175

gtg?ttc?ggc?acc?tgg?ctg?ccg?cac?cgc?ccc?ggc?cac?gac?gcg?ttc?ccg???????576

Val?Phe?Gly?Thr?Trp?Leu?Pro?His?Arg?Pro?Gly?His?Asp?Ala?Phe?Pro

180?????????????????185?????????????????190

gac?cgc?cac?aat?gcg?cgg?tcg?tcg?cgg?atc?agc?gac?ccc?gtg?tcg?ctg???????624

Asp?Arg?His?Asn?Ala?Arg?Ser?Ser?Arg?Ile?Ser?Asp?Pro?Val?Ser?Leu

195?????????????????200?????????????????205

ctg?acc?tgc?ttt?cac?ttt?ggc?ggt?tat?cat?cac?gaa?cac?cac?ctg?cac???????672

Leu?Thr?Cys?Phe?His?Phe?Gly?Gly?Tyr?His?His?Glu?His?His?Leu?His

210?????????????????215?????????????????220

ccg?acg?gtg?ccg?tgg?tgg?cgc?ctg?ccc?agc?acc?cgc?acc?aag?ggg?gac???????720

Pro?Thr?Val?Pro?Trp?Trp?Arg?Leu?Pro?Ser?Thr?Arg?Thr?Lys?Gly?Asp

225?????????????????230?????????????????235?????????????????240

acc?gca?tga???????????????????????????????????????????????????????????729

Thr?Ala

<210>6

<211>242

<212>PRT

＜213〉orange Agrobacterium

<400>6

Met?Ser?Ala?His?Ala?Leu?Pro?Lys?Ala?Asp?Leu?Thr?Ala?Thr?Ser?Leu

1???????????????5???????????????????10??????????????????15

Ile?Val?Ser?Gly?Gly?Ile?Ile?Ala?Ala?Trp?Leu?Ala?Leu?His?Val?His

20??????????????????25??????????????????30

Ala?Leu?Trp?Phe?Leu?Asp?Ala?Ala?Ala?His?Pro?Ile?Leu?Ala?Ile?Ala

35??????????????????40??????????????????45

Asn?Phe?Leu?Gly?Leu?Thr?Trp?Leu?Ser?Val?Gly?Leu?Phe?Ile?Ile?Ala

50??????????????????55??????????????????60

His?Asp?Ala?Met?His?Gly?Ser?Val?Val?Pro?Gly?Arg?Pro?Arg?Ala?Asn

65??????????????????70??????????????????75??????????????????80

Ala?Ala?Met?Gly?Gln?Leu?Val?Leu?Trp?Leu?Tyr?Ala?Gly?Phe?Ser?Trp

85??????????????????90??????????????????95

Arg?Lys?Met?Ile?Val?Lys?His?Met?Ala?His?His?Arg?His?Ala?Gly?Thr

100?????????????????105?????????????????110

Asp?Asp?Asp?Pro?Asp?Phe?Asp?His?Gly?Gly?Pro?Val?Arg?Trp?Tyr?Ala

115?????????????????120?????????????????125

Arg?Phe?Ile?Gly?Thr?Tyr?Phe?Gly?Trp?Arg?Glu?Gly?Leu?Leu?Leu?Pro

130?????????????????135?????????????????140

Val?Ile?Val?Thr?Val?Tyr?Ala?Leu?Ile?Leu?Gly?Asp?Arg?Trp?Met?Tyr

145?????????????????150?????????????????155?????????????????160

Val?Val?Phe?Trp?Pro?Leu?Pro?Ser?Ile?Leu?Ala?Ser?Ile?Gln?Leu?Phe

165?????????????????170?????????????????175

Val?Phe?Gly?Thr?Trp?Leu?Pro?His?Arg?Pro?Gly?His?Asp?Ala?Phe?Pro

180?????????????????185?????????????????190

Asp?Arg?His?Asn?Ala?Arg?Ser?Ser?Arg?Ile?Ser?Asp?Pro?Val?Ser?Leu

195?????????????????200?????????????????205

Leu?Thr?Cys?Phe?His?Phe?Gly?Gly?Tyr?His?His?Glu?His?His?Leu?His

210?????????????????215?????????????????220

Pro?Thr?Val?Pro?Trp?Trp?Arg?Leu?Pro?Ser?Thr?Arg?Thr?Lys?Gly?Asp

225?????????????????230?????????????????235?????????????????240

Thr?Ala

<210>7

<211>1631

<212>DNA

＜213〉Alcaligenes (Alcaligenes sp.)

<220>

<221>CDS

<222>(99)..(827)

<223>

<400>7

ctgcaggccg?ggcccggtgg?ccaatggtcg?caaccggcag?gactggaaca?ggacggcggg??????60

ccggtctagg?ctgtcgccct?acgcagcagg?agtttcgg?atg?tcc?gga?cgg?aag?cct?????116

Met?Ser?Gly?Arg?Lys?Pro

1???????????????5

ggc?aca?act?ggc?gac?acg?atc?gtc?aat?ctc?ggt?ctg?acc?gcc?gcg?atc???????164

Gly?Thr?Thr?Gly?Asp?Thr?Ile?Val?Asn?Leu?Gly?Leu?Thr?Ala?Ala?Ile

10??????????????????15??????????????????20

ctg?ctg?tgc?tgg?ctg?gtc?ctg?cac?gcc?ttt?acg?cta?tgg?ttg?cta?gat???????212

Leu?Leu?Cys?Trp?Leu?Val?Leu?His?Ala?Phe?Thr?Leu?Trp?Leu?Leu?Asp

25??????????????????30??????????????????35

gcg?gcc?gcg?cat?ccg?ctg?ctt?gcc?gtg?ctg?tgc?ctg?gct?ggg?ctg?acc???????260

Ala?Ala?Ala?His?Pro?Leu?Leu?Ala?Val?Leu?Cys?Leu?Ala?Gly?Leu?Thr

40??????????????????45??????????????????50

tgg?ctg?tcg?gtc?ggg?ctg?ttc?atc?atc?gcg?cat?gac?gca?atg?cac?ggg???????308

Trp?Leu?Ser?Val?Gly?Leu?Phe?Ile?Ile?Ala?His?Asp?Ala?Met?His?Gly

55??????????????????60??????????????????65??????????????????70

tcc?gtg?gtg?ccg?ggg?cgg?ccg?cgc?gcc?aat?gcg?gcg?atc?ggg?caa?ctg???????356

Ser?Val?Val?Pro?Gly?Arg?Pro?Arg?Ala?Asn?Ala?Ala?Ile?Gly?Gln?Leu

75??????????????????80??????????????????85

gcg?ctg?tgg?ctc?tat?gcg?ggg?ttc?tcg?tgg?ccc?aag?ctg?atc?gcc?aag???????404

Ala?Leu?Trp?Leu?Tyr?Ala?Gly?Phe?Ser?Trp?Pro?Lys?Leu?Ile?Ala?Lys

90??????????????????95??????????????????100

cac?atg?acg?cat?cac?cgg?cac?gcc?ggc?acc?gac?aac?gat?ccc?gat?ttc???????452

His?Met?Thr?His?His?Arg?His?Ala?Gly?Thr?Asp?Asn?Asp?Pro?Asp?Phe

105?????????????????110?????????????????115

ggt?cac?gga?ggg?ccc?gtg?cgc?tgg?tac?ggc?agc?ttc?gtc?tcc?acc?tat???????500

Gly?His?Gly?Gly?Pro?Val?Arg?Trp?Tyr?Gly?Ser?Phe?Val?Ser?Thr?Tyr

120?????????????????125?????????????????130

ttc?ggc?tgg?cga?gag?gga?ctg?ctg?cta?ccg?gtg?atc?gtc?acc?acc?tat???????548

Phe?Gly?Trp?Arg?Glu?Gly?Leu?Leu?Leu?Pro?Val?Ile?Val?Thr?Thr?Tyr

135?????????????????140?????????????????145?????????????????150

gcg?ctg?atc?ctg?ggc?gat?cgc?tgg?atg?tat?gtc?atc?ttc?tgg?ccg?gtc???????596

Ala?Leu?Ile?Leu?Gly?Asp?Arg?Trp?Met?Tyr?Val?Ile?Phe?Trp?Pro?Val

155?????????????????160?????????????????165

ccg?gcc?gtt?ctg?gcg?tcg?atc?cag?att?ttc?gtc?ttc?gga?act?tgg?ctg???????644

Pro?Ala?Val?Leu?Ala?Ser?Ile?Gln?Ile?Phe?Val?Phe?Gly?Thr?Trp?Leu

170?????????????????175?????????????????180

ccc?cac?cgc?ccg?gga?cat?gac?gat?ttt?ccc?gac?cgg?cac?aac?gcg?agg???????692

Pro?His?Arg?Pro?Gly?His?Asp?Asp?Phe?Pro?Asp?Arg?His?Asn?Ala?Arg

185?????????????????190?????????????????195

tcg?acc?ggc?atc?ggc?gac?ccg?ttg?tca?cta?ctg?acc?tgc?ttc?cat?ttc???????740

Ser?Thr?Gly?Ile?Gly?Asp?Pro?Leu?Ser?Leu?Leu?Thr?Cys?Phe?His?Phe

200?????????????????205?????????????????210

ggc?ggc?tat?cac?cac?gaa?cat?cac?ctg?cat?ccg?cat?gtg?ccg?tgg?tgg???????788

Gly?Gly?Tyr?His?His?Glu?His?His?Leu?His?Pro?His?Val?Pro?Trp?Trp

215?????????????????220?????????????????225?????????????????230

cgc?ctg?cct?cgt?aca?cgc?aag?acc?gga?ggc?cgc?gca?tga?cgcaattcct????????837

Arg?Leu?Pro?Arg?Thr?Arg?Lys?Thr?Gly?Gly?Arg?Ala

235?????????????????240

cattgtcgtg?gcgacagtcc?tcgtgatgga?gctgaccgcc?tattccgtcc?accgctggat?????897

tatgcacggc?cccctaggct?ggggctggca?caagtcccat?cacgaagagc?acgaccacgc?????957

gttggagaag?aacgacotct?acggcgtcgt?cttcgcggtg?ctggcgacga?tcctcttcac????1017

cgtgggcgcc?tattggtggc?cggtgctgtg?gtggatcgcc?ctgggcatga?cggtctatgg????1077

gttgatctat?ttcatcctgc?acgacgggct?tgtgcatcaa?cgctggccgt?ttcggtatat????1137

tccgcggcgg?ggctatttcc?gcaggctcta?ccaagctcat?cgcctgcacc?acgcggtcga????1197

ggggcgggac?cactgcgtca?gcttcggctt?catctatgcc?ccacccgtgg?acaagctgaa????1257

gcaggatctg?aagcggtcgg?gtgtcctgcg?cccccaggac?gagcgtccgt?cgtgatctct????1317

gatcccggcg?tggccgcatg?aaatccgacg?tgctgctggc?aggggccggc?cttgccaacg????1377

gactgatcgc?gctggcgatc?cgcaaggcgc?ggcccgacct?tcgcgtgctg?ctgctggacc????1437

gtgcggcggg?cgcctcggac?gggcatactt?ggtcctgcca?cgacaccgat?ttggcgccgc????1497

actggctgga?ccgcctgaag?ccgatcaggc?gtggcgactg?gcccgatcag?gaggtgcggt????1557

tcccagacca?ttcgcgaagg?ctccgggccg?gatatggctc?gatcgacggg?cgggggctga????1617

tgcgtgcggt?gacc??????????????????????????????????????????????????????1631

<210>8

<211>242

<212>PRT

＜213〉Alcaligenes

<400>8

Met?Ser?Gly?Arg?Lys?Pro?Gly?Thr?Thr?Gly?Asp?Thr?Ile?Val?Asn?Leu

1???????????????5???????????????????10??????????????????15

Gly?Leu?Thr?Ala?Ala?Ile?Leu?Leu?Cys?Trp?Leu?Val?Leu?His?Ala?Phe

20??????????????????25??????????????????30

Thr?Leu?Trp?Leu?Leu?Asp?Ala?Ala?Ala?His?Pro?Leu?Leu?Ala?Val?Leu

35??????????????????40??????????????????45

Cys?Leu?Ala?Gly?Leu?Thr?Trp?Leu?Ser?Val?Gly?Leu?Phe?Ile?Ile?Ala

50??????????????????55??????????????????60

His?Asp?Ala?Met?His?Gly?Ser?Val?Val?Pro?Gly?Arg?Pro?Arg?Ala?Asn

65??????????????????70??????????????????75??????????????????80

Ala?Ala?Ile?Gly?Gln?Leu?Ala?Leu?Trp?Leu?Tyr?Ala?Gly?Phe?Ser?Trp

85??????????????????90??????????????????95

Pro?Lys?Leu?Ile?Ala?Lys?His?Met?Thr?His?His?Arg?His?Ala?Gly?Thr

100?????????????????105?????????????????110

Asp?Asn?Asp?Pro?Asp?Phe?Gly?His?Gly?Gly?Pro?Val?Arg?Trp?Tyr?Gly

115?????????????????120?????????????????125

Ser?Phe?Val?Ser?Thr?Tyr?Phe?Gly?Trp?Arg?Glu?Gly?Leu?Leu?Leu?Pro

130?????????????????135?????????????????140

Val?Ile?Val?Thr?Thr?Tyr?Ala?Leu?Ile?Leu?Gly?Asp?Arg?Trp?Met?Tyr

145?????????????????150?????????????????155?????????????????160

Val?Ile?Phe?Trp?Pro?Val?Pro?Ala?Val?Leu?Ala?Ser?Ile?Gln?Ile?Phe

165?????????????????170?????????????????175

Val?Phe?Gly?Thr?Trp?Leu?Pro?His?Arg?Pro?Gly?His?Asp?Asp?Phe?Pro

180?????????????????185?????????????????190

Asp?Arg?His?Asn?Ala?Arg?Ser?Thr?Gly?Ile?Gly?Asp?Pro?Leu?Ser?Leu

195?????????????????200?????????????????205

Leu?Thr?Cys?Phe?His?Phe?Gly?Gly?Tyr?His?His?Glu?His?His?Leu?His

210?????????????????215?????????????????220

Pro?His?Val?Pro?Trp?Trp?Arg?Leu?Pro?Arg?Thr?Arg?Lys?Thr?Gly?Gly

225?????????????????230?????????????????235?????????????????240

Arg?Ala

<210>9

<211>729

<212>DNA

<213>Paracoccus?marcusii

<220>

<221>CDS

<222>(1)..(729)

<223>

<400>9

atg?agc?gca?cat?gcc?ctg?ccc?aag?gca?gat?ctg?acc?gcc?aca?agc?ctg????????48

Met?Ser?Ala?His?Ala?Leu?Pro?Lys?Ala?Asp?Leu?Thr?Ala?Thr?Ser?Leu

1???????????????5???????????????????10??????????????????15

atc?gtc?tcg?ggc?ggc?atc?arc?gcc?gca?tgg?ctg?gcc?ctg?cat?gtg?cat????????96

Ile?Val?Ser?Gly?Gly?Ile?Ile?Ala?Ala?Trp?Leu?Ala?Leu?His?Val?His

20??????????????????25??????????????????30

gcg?ctg?tgg?ttt?ctg?gac?gcg?gcg?gcc?cat?ccc?atc?ctg?gcg?gtc?gcg???????144

Ala?Leu?Trp?Phe?Leu?Asp?Ala?Ala?Ala?His?Pro?Ile?Leu?Ala?Val?Ala

35??????????????????40??????????????????45

aat?ttc?ctg?ggg?ctg?acc?tgg?ctg?tcg?gtc?gga?ttg?ttc?atc?atc?gcg???????192

Asn?Phe?Leu?Gly?Leu?Thr?Trp?Leu?Ser?Val?Gly?Leu?Phe?Ile?Ile?Ala

50??????????????????55??????????????????60

cat?gac?gcg?atg?cac?ggg?tcg?gtc?gtg?ccg?ggg?cgt?ccg?cgc?gcc?aat???????240

His?Asp?Ala?Met?His?Gly?Ser?Val?Val?Pro?Gly?Arg?Pro?Arg?Ala?Asn

65??????????????????70??????????????????75??????????????????80

gcg?gcg?atg?ggc?cag?ctt?gtc?ctg?tgg?ctg?tat?gcc?gga?ttt?tcg?tgg???????288

Ala?Ala?Met?Gly?Gln?Leu?Val?Leu?Trp?Leu?Tyr?Ala?Gly?Phe?Ser?Trp

85??????????????????90??????????????????95

cgc?aag?atg?atc?gtc?aag?cac?atg?gcc?cat?cac?cgc?cat?gcc?gga?acc???????336

Arg?Lys?Met?Ile?Val?Lys?His?Met?Ala?His?His?Arg?His?Ala?Gly?Thr

100?????????????????105?????????????????110

gac?gac?gac?cca?gat?ttc?gac?cat?ggc?ggc?ccg?gtc?cgc?tgg?tac?gcc???????384

Asp?Asp?Asp?Pro?Asp?Phe?Asp?His?Gly?Gly?Pro?Val?Arg?Trp?Tyr?Ala

115?????????????????120?????????????????125

cgc?ttc?atc?ggc?acc?tat?ttc?ggc?tgg?cgc?gag?ggg?ctg?ctg?ctg?ccc???????432

Arg?Phe?Ile?Gly?Thr?Tyr?Phe?Gly?Trp?Arg?Glu?Gly?Leu?Leu?Leu?Pro

130?????????????????135?????????????????140

gtc?atc?gtg?acg?gtc?tat?gcg?ctg?atc?ctg?ggg?gat?cgc?tgg?atg?tac???????480

Val?Ile?Val?Thr?Val?Tyr?Ala?Leu?Ile?Leu?Gly?Asp?Arg?Trp?Met?Tyr

145?????????????????150?????????????????155?????????????????160

gtg?gtc?ttc?tgg?ccg?ttg?ccg?tcg?atc?ctg?gcg?tcg?atc?cag?ctg?ttc???????528

Val?Val?Phe?Trp?Pro?Leu?Pro?Ser?Ile?Leu?Ala?Ser?Ile?Gln?Leu?Phe

165?????????????????170?????????????????175

gtg?ttc?ggc?act?tgg?ctg?ccg?cac?cgc?ccc?ggc?cac?gac?gcg?ttc?ccg???????576

Val?Phe?Gly?Thr?Trp?Leu?Pro?His?Arg?Pro?Gly?His?Asp?Ala?Phe?Pro

180?????????????????185??????????????????190

gac?cgc?cat?aat?gcg?cgg?tcg?tcg?cgg?atc?agc?gac?cct?gtg?tcg?ctg???????624

Asp?Arg?His?Asn?Ala?Arg?Ser?Ser?Arg?Ile?Ser?Asp?Pro?Val?Ser?Leu

195?????????????????200?????????????????205

ctg?acc?tgc?ttt?cat?ttt?ggc?ggt?tat?cat?cac?gaa?cac?cac?ctg?cac???????672

Leu?Thr?Cys?Phe?His?Phe?Gly?Gly?Tyr?His?His?Glu?His?His?Leu?His

210?????????????????215?????????????????220

ccg?acg?gtg?ccg?tgg?tgg?cgc?ctg?ccc?agc?acc?cgc?acc?aag?ggg?gac??????720

Pro?Thr?Val?Pro?Trp?Trp?Arg?Leu?Pro?Ser?Thr?Arg?Thr?Lys?Gly?Asp

225?????????????????230?????????????????235?????????????????240

acc?gca?tga??????????????????????????????????????????????????????????729

Thr?Ala

<210>10

<211>242

<212>PRT

<213>Paracoccus?marcusii

<400>10

Met?Ser?Ala?His?Ala?Leu?Pro?Lys?Ala?Asp?Leu?Thr?Ala?Thr?Ser?Leu

1???????????????5???????????????????10??????????????????15

Ile?Val?Ser?Gly?Gly?Ile?Ile?Ala?Ala?Trp?Leu?Ala?Leu?His?Val?His

20??????????????????25??????????????????30

Ala?Leu?Trp?Phe?Leu?Asp?Ala?Ala?Ala?His?Pro?Ile?Leu?Ala?Val?Ala

35??????????????????40??????????????????45

Asn?Phe?Leu?Gly?Leu?Thr?Trp?Leu?Ser?Val?Gly?Leu?Phe?Ile?Ile?Ala

50??????????????????55??????????????????60

His?Asp?Ala?Met?His?Gly?Ser?Val?Val?Pro?Gly?Arg?Pro?Arg?Ala?Asn

65??????????????????70??????????????????75??????????????????80

Ala?Ala?Met?Gly?Gln?Leu?Val?Leu?Trp?Leu?Tyr?Ala?Gly?Phe?Ser?Trp

85??????????????????90???????????????????95

Arg?Lys?Met?Ile?Val?Lys?His?Met?Ala?His?His?Arg?His?Ala?Gly?Thr

100?????????????????105?????????????????110

Asp?Asp?Asp?Pro?Asp?Phe?Asp?His?Gly?Gly?Pro?Val?Arg?Trp?Tyr?Ala

115??????????????????120?????????????????125

Arg?Phe?Ile?Gly?Thr?Tyr?Phe?Gly?Trp?Arg?Glu?Gly?Leu?Leu?Leu?Pro

130?????????????????135?????????????????140

Val?Ile?Val?Thr?Val?Tyr?Ala?Leu?Ile?Leu?Gly?Asp?Arg?Trp?Met?Tyr

145?????????????????150?????????????????155?????????????????160

Val?Val?Phe?Trp?Pro?Leu?Pro?Ser?Ile?Leu?Ala?Ser?Ile?Gln?Leu?Phe

165?????????????????170?????????????????175

Val?Phe?Gly?Thr?Trp?Leu?Pro?His?Arg?Pro?Gly?His?Asp?Ala?Phe?Pro

180?????????????????185?????????????????190

Asp?Arg?His?Asn?Ala?Arg?Ser?Ser?Arg?Ile?Ser?Asp?Pro?Val?Ser?Leu

195??????????????????200?????????????????205

Leu?Thr?Cys?Phe?His?Phe?Gly?Gly?Tyr?His?His?Glu?His?His?Leu?His

210?????????????????215?????????????????220

Pro?Thr?Val?Pro?Trp?Trp?Arg?Leu?Pro?Ser?Thr?Arg?Thr?Lys?Gly?Asp

225?????????????????230?????????????????235?????????????????240

Thr?Ala

<210>11

<211>1629

<212>DNA

＜213〉Synechococcus (Synechococcus sp.)

<220>

<221>CDS

<222>(1)..(1629)

<223>

<400>11

atg?atc?acc?acc?gat?gtt?gtc?att?att?ggg?gcg?ggg?cac?aat?ggc?tta????????48

Met?Ile?Thr?Thr?Asp?Val?Val?Ile?Ile?Gly?Ala?Gly?His?Asn?Gly?Leu

1???????????????5???????????????????10??????????????????15

gtc?tgt?gca?gcc?tat?ttg?ctc?caa?cgg?ggc?ttg?ggg?gtg?acg?tta?cta????????96

Val?Cys?Ala?Ala?Tyr?Leu?Leu?Gln?Arg?Gly?Leu?Gly?Val?Thr?Leu?Leu

20??????????????????25??????????????????30

gaa?aag?cgg?gaa?gta?cca?ggg?ggg?gcg?gcc?acc?aca?gaa?gct?ctc?atg???????144

Glu?Lys?Arg?Glu?Val?Pro?Gly?Gly?Ala?Ala?Thr?Thr?Glu?Ala?Leu?Met

35??????????????????40??????????????????45

ccg?gag?cta?tcc?ccc?cag?ttt?cgc?ttt?aac?cgc?tgt?gcc?att?gac?cac???????192

Pro?Glu?Leu?Ser?Pro?Gln?Phe?Arg?Phe?Asn?Arg?Cys?Ala?Ile?Asp?His

50??????????????????55??????????????????60

gaa?ttt?atc?ttt?ctg?ggg?ccg?gtg?ttg?cag?gag?cta?aat?tta?gcc?cag???????240

Glu?Phe?Ile?Phe?Leu?Gly?Pro?Val?Leu?Gln?Glu?Leu?Asn?Leu?Ala?Gln

65??????????????????70???????????????????75?????????????????80

tat?ggt?ttg?gaa?tat?tta?ttt?tgt?gac?ccc?agt?gtt?ttt?tgt?ccg?ggg???????288

Tyr?Gly?Leu?Glu?Tyr?Leu?Phe?Cys?Asp?Pro?Ser?Val?Phe?Cys?Pro?Gly

85??????????????????90??????????????????95

ctg?gat?ggc?caa?gct?ttt?atg?agc?tac?cgt?tcc?cta?gaa?aaa?acc?tgt???????336

Leu?Asp?Gly?Gln?Ala?Phe?Met?Ser?Tyr?Arg?Ser?Leu?Glu?Lys?Thr?Cys

100?????????????????105?????????????????110

gcc?cac?att?gcc?acc?tat?agc?ccc?cga?gat?gcg?gaa?aaa?tat?cgg?caa???????384

Ala?His?Ile?Ala?Thr?Tyr?Ser?Pro?Arg?Asp?Ala?Glu?Lys?Tyr?Arg?Gln

115?????????????????120?????????????????125

ttt?gtc?aat?tat?tgg?acg?gat?ttg?ctc?aac?gct?gtc?cag?cct?gct?ttt???????432

Phe?Val?Asn?Tyr?Trp?Thr?Asp?Leu?Leu?Asn?Ala?Val?Gln?Pro?Ala?Phe

130?????????????????135?????????????????140

aat?gct?ccg?ccc?cag?gct?tta?cta?gat?tta?gcc?ctg?aac?tat?ggt?tgg???????480

Asn?Ala?Pro?Pro?Gln?Ala?Leu?Leu?Asp?Leu?Ala?Leu?Asn?Tyr?Gly?Trp

145????????????????150??????????????????155?????????????????160

gaa?aac?tta?aaa?tcc?gtg?ctg?gcg?atc?gcc?ggg?tcg?aaa?acc?aag?gcg???????528

Glu?Asn?Leu?Lys?Ser?Val?Leu?Ala?Ile?Ala?Gly?Ser?Lys?Thr?Lys?Ala

165?????????????????170?????????????????175

ttg?gat?ttt?atc?cgc?act?atg?atc?ggc?tcc?ccg?gaa?gat?gtg?ctc?aat???????576

Leu?Asp?Phe?Ile?Arg?Thr?Met?Ile?Gly?Ser?Pro?Glu?Asp?Val?Leu?Asn

180?????????????????185?????????????????190

gaa?tgg?ttc?gac?agc?gaa?cgg?gtt?aaa?gct?cct?tta?gct?aga?cta?tgt???????624

Glu?Trp?Phe?Asp?Ser?Glu?Arg?Val?Lys?Ala?Pro?Leu?Ala?Arg?Leu?Cys

195?????????????????200?????????????????205

tcg?gaa?att?ggc?gct?ccc?cca?tcc?caa?aag?ggt?agt?agc?tcc?ggc?atg???????672

Ser?Glu?Ile?Gly?Ala?Pro?Pro?Ser?Gln?Lys?Gly?Ser?Ser?Ser?Gly?Met

210?????????????????215?????????????????220

atg?atg?gtg?gcc?atg?cgg?cat?ttg?gag?gga?att?gcc?aga?cca?aaa?gga???????720

Met?Met?Val?Ala?Met?Arg?His?Leu?Glu?Gly?Ile?Ala?Arg?Pro?Lys?Gly

225?????????????????230?????????????????235?????????????????240

ggc?act?gga?gcc?ctc?aca?gaa?gcc?ttg?gtg?aag?tta?gtg?caa?gcc?caa???????768

Gly?Thr?Gly?Ala?Leu?Thr?Glu?Ala?Leu?Val?Lys?Leu?Val?Gln?Ala?Gln

245?????????????????250?????????????????255

ggg?gga?aaa?atc?ctc?act?gac?caa?acc?gtc?aaa?cgg?gta?ttg?gtg?gaa???????816

Gly?Gly?Lys?Ile?Leu?Thr?Asp?Gln?Thr?Val?Lys?Arg?Val?Leu?Val?Glu

260?????????????????265?????????????????270

aac?aac?cag?gcg?atc?ggg?gtg?gag?gta?gct?aac?gga?gaa?cag?tac?cgg???????864

Asn?Asn?Gln?Ala?Ile?Gly?Val?Glu?Val?Ala?Asn?Gly?Glu?Gln?Tyr?Arg

275?????????????????280?????????????????285

gcc?aaa?aaa?ggc?gtg?att?tct?aac?atc?gat?gcc?cgc?cgt?tta?ttt?ttg???????912

Ala?Lys?Lys?Gly?Val?Ile?Ser?Asn?Ile?Asp?Ala?Arg?Arg?Leu?Phe?Leu

290?????????????????295?????????????????????300

caa?ttg?gtg?gaa?ccg?ggg?gcc?cta?gcc?aag?gtg?aat?caa?aac?cta?ggg???????960

Gln?Leu?Val?Glu?Pro?Gly?Ala?Leu?Ala?Lys?Val?Asn?Gln?Asn?Leu?Gly

305?????????????????310?????????????????315?????????????????320

gaa?cga?ctg?gaa?cgg?cgc?act?gtg?aac?aat?aac?gaa?gcc?att?tta?aaa??????1008

Glu?Arg?Leu?Glu?Arg?Arg?Thr?Val?Asn?Asn?Asn?Glu?Ala?Ile?Leu?Lys

325?????????????????330?????????????????335

atc?gat?tgt?gcc?ctc?tcc?ggt?tta?ccc?cac?ttc?act?gcc?atg?gcc?ggg??????1056

Ile?Asp?Cys?Ala?Leu?Ser?Gly?Leu?Pro?His?Phe?Thr?Ala?Met?Ala?Gly

340?????????????????345?????????????????350

ccg?gag?gat?cta?acg?gga?act?att?ttg?att?gcc?gac?tcg?gta?cgc?cat??????1104

Pro?Glu?Asp?Leu?Thr?Gly?Thr?Ile?Leu?Ile?Ala?Asp?Ser?Val?Arg?His

355?????????????????360?????????????????365

gtc?gag?gaa?gcc?cac?gcc?ctc?att?gcc?ttg?ggg?caa?att?ccc?gat?gct??????1152

Val?Glu?Glu?Ala?His?Ala?Leu?Ile?Ala?Leu?Gly?Gln?Ile?Pro?Asp?Ala

370?????????????????375?????????????????380

aat?ccg?tct?tta?tat?ttg?gat?att?ccc?act?gta?ttg?gac?ccc?acc?atg??????1200

Asn?Pro?Ser?Leu?Tyr?Leu?Asp?Ile?Pro?Thr?Val?Leu?Asp?Pro?Thr?Met

385?????????????????390?????????????????395?????????????????400

gcc?ccc?cct?ggg?cag?cac?acc?ctc?tgg?atc?gaa?ttt?ttt?gcc?ccc?tac??????1248

Ala?Pro?Pro?Gly?Gln?His?Thr?Leu?Trp?Ile?Glu?Phe?Phe?Ala?Pro?Tyr

405?????????????????410?????????????????415

cgc?atc?gcc?ggg?ttg?gaa?ggg?aca?ggg?tta?atg?ggc?aca?ggt?tgg?acc??????1296

Arg?Ile?Ala?Gly?Leu?Glu?Gly?Thr?Gly?Leu?Met?Gly?Thr?Gly?Trp?Thr

420?????????????????425?????????????????430

gat?gag?tta?aag?gaa?aaa?gtg?gcg?gat?cgg?gtg?att?gat?aaa?tta?acg??????1344

Asp?Glu?Leu?Lys?Glu?Lys?Val?Ala?Asp?Arg?Val?Ile?Asp?Lys?Leu?Thr

435?????????????????440?????????????????445

gac?tat?gcc?cct?aac?cta?aaa?tct?ctg?atc?att?ggt?cgc?cga?gtg?gaa??????1392

Asp?Tyr?Ala?Pro?Asn?Leu?Lys?Ser?Leu?Ile?Ile?Gly?Arg?Arg?Val?Glu

450?????????????????455?????????????????460

agt?ccc?gcc?gaa?ctg?gcc?caa?cgg?ctg?gga?agt?tac?aac?ggc?aat?gtc??????1440

Ser?Pro?Ala?Glu?Leu?Ala?Gln?Arg?Leu?Gly?Ser?Tyr?Asn?Gly?Asn?Val

465?????????????????470?????????????????475?????????????????480

tat?cat?ctg?gat?atg?agt?ttg?gac?caa?atg?atg?ttc?ctc?cgg?cct?cta??????1488

Tyr?His?Leu?Asp?Met?Ser?Leu?Asp?Gln?Met?Met?Phe?Leu?Arg?Pro?Leu

485?????????????????490?????????????????495

ccg?gaa?att?gcc?aac?tac?caa?acc?ccc?atc?aaa?aat?ctt?tac?tta?aca??????1536

Pro?Glu?Ile?Ala?Asn?Tyr?Gln?Thr?Pro?Ile?Lys?Asn?Leu?Tyr?Leu?Thr

500?????????????????505?????????????????510

ggg?gcg?ggt?acc?cat?ccc?ggt?ggc?tcc?ata?tca?ggt?atg?ccc?ggt?aga??????1584

Gly?Ala?Gly?Thr?His?Pro?Gly?Gly?Ser?Ile?Ser?Gly?Met?Pro?Gly?Arg

515?????????????????520?????????????????525

aat?tgc?gct?cgg?gtc?ttt?tta?aaa?caa?caa?cgt?cgt?ttt?tgg?taa??????????1629

Asn?Cys?Ala?Arg?Val?Phe?Leu?Lys?Gln?Gln?Arg?Arg?Phe?Trp

530?????????????????535?????????????????540

<210>12

<211>542

<212>PRT

＜213〉Synechococcus

<400>12

Met?Ile?Thr?Thr?Asp?Val?Val?Ile?Ile?Gly?Ala?Gly?His?Asn?Gly?Leu

1???????????????5???????????????????10??????????????????15

Val?Cys?Ala?Ala?Tyr?Leu?Leu?Gln?Arg?Gly?Leu?Gly?Val?Thr?Leu?Leu

20??????????????????25??????????????????30

Glu?Lys?Arg?Glu?Val?Pro?Gly?Gly?Ala?Ala?Thr?Thr?Glu?Ala?Leu?Met

35??????????????????40??????????????????45

Pro?Glu?Leu?Ser?Pro?Gln?Phe?Arg?Phe?Asn?Arg?Cys?Ala?Ile?Asp?His

50??????????????????55??????????????????60

Glu?Phe?Ile?Phe?Leu?Gly?Pro?Val?Leu?Gln?Glu?Leu?Asn?Leu?Ala?Gln

65??????????????????70??????????????????75??????????????????80

Tyr?Gly?Leu?Glu?Tyr?Leu?Phe?Cys?Asp?Pro?Ser?Val?Phe?Cys?Pro?Gly

85??????????????????90??????????????????95

Leu?Asp?Gly?Gln?Ala?Phe?Met?Ser?Tyr?Arg?Ser?Leu?Glu?Lys?Thr?Cys

100?????????????????105?????????????????110

Ala?His?Ile?Ala?Thr?Tyr?Ser?Pro?Arg?Asp?Ala?Glu?Lys?Tyr?Arg?Gln

115?????????????????120?????????????????125

Phe?Val?Asn?Tyr?Trp?Thr?Asp?Leu?Leu?Asn?Ala?Val?Gln?Pro?Ala?Phe

130?????????????????135?????????????????140

Asn?Ala?Pro?Pro?Gln?Ala?Leu?Leu?Asp?Leu?Ala?Leu?Asn?Tyr?Gly?Trp

145?????????????????150?????????????????155?????????????????160

Glu?Asn?Leu?Lys?Ser?Val?Leu?Ala?Ile?Ala?Gly?Ser?Lys?Thr?Lys?Ala

165?????????????????170?????????????????175

Leu?Asp?Phe?Ile?Arg?Thr?Met?Ile?Gly?Ser?Pro?Glu?Asp?Val?Leu?Asn

180?????????????????185?????????????????190

Glu?Trp?Phe?Asp?Ser?Glu?Arg?Val?Lys?Ala?Pro?Leu?Ala?Arg?Leu?Cys

195?????????????????200?????????????????205

Ser?Glu?Ile?Gly?Ala?Pro?Pro?Ser?Gln?Lys?Gly?Ser?Ser?Ser?Gly?Met

210?????????????????215?????????????????220

Met?Met?Val?Ala?Met?Arg?His?Leu?Glu?Gly?Ile?Ala?Arg?Pro?Lys?Gly

225?????????????????230?????????????????235?????????????????240

Gly?Thr?Gly?Ala?Leu?Thr?Glu?Ala?Leu?Val?Lys?Leu?Val?Gln?Ala?Gln

245?????????????????250?????????????????255

Gly?Gly?Lys?Ile?Leu?Thr?Asp?Gln?Thr?Val?Lys?Arg?Val?Leu?Val?Glu

260?????????????????265?????????????????270

Asn?Asn?Gln?Ala?Ile?Gly?Val?Glu?Val?Ala?Asn?Gly?Glu?Gln?Tyr?Arg

275?????????????????280?????????????????285

Ala?Lys?Lys?Gly?Val?Ile?Ser?Asn?Ile?Asp?Ala?Arg?Arg?Leu?Phe?Leu

290?????????????????295?????????????????300

Gln?Leu?Val?Glu?Pro?Gly?Ala?Leu?Ala?Lys?Val?Asn?Gln?Asn?Leu?Gly

305?????????????????310?????????????????315?????????????????320

Glu?Arg?Leu?Glu?Arg?Arg?Thr?Val?Asn?Asn?Asn?Glu?Ala?Ile?Leu?Lys

325?????????????????330?????????????????335

Ile?Asp?Cys?Ala?Leu?Ser?Gly?Leu?Pro?His?Phe?Thr?Ala?Met?Ala?Gly

340?????????????????345?????????????????350

Pro?Glu?Asp?Leu?Thr?Gly?Thr?Ile?Leu?Ile?Ala?Asp?Ser?Val?Arg?His

355?????????????????360?????????????????365

Val?Glu?Glu?Ala?His?Ala?Leu?Ile?Ala?Leu?Gly?Gln?Ile?Pro?Asp?Ala

370?????????????????375?????????????????380

Asn?Pro?Ser?Leu?Tyr?Leu?Asp?Ile?Pro?Thr?Val?Leu?Asp?Pro?Thr?Met

385?????????????????390?????????????????395?????????????????400

Ala?Pro?Pro?Gly?Gln?His?Thr?Leu?Trp?Ile?Glu?Phe?Phe?Ala?Pro?Tyr

405?????????????????410?????????????????415

Arg?Ile?Ala?Gly?Leu?Glu?Gly?Thr?Gly?Leu?Met?Gly?Thr?G1y?Trp?Thr

420?????????????????425?????????????????430

Asp?Glu?Leu?Lys?Glu?Lys?Val?Ala?Asp?Arg?Val?Ile?Asp?Lys?Leu?Thr

435?????????????????440?????????????????445

Asp?Tyr?Ala?Pro?Asn?Leu?Lys?Ser?Leu?Ile?Ile?Gly?Arg?Arg?Val?Glu

450?????????????????455?????????????????460

Ser?Pro?Ala?Glu?Leu?Ala?Gln?Arg?Leu?Gly?Ser?Tyr?Asn?Gly?Asn?Val

465?????????????????470?????????????????475?????????????????480

Tyr?His?Leu?Asp?Met?Ser?Leu?Asp?Gln?Met?Met?Phe?Leu?Arg?Pro?Leu

485?????????????????490?????????????????495

Pro?Glu?Ile?Ala?Asn?Tyr?Gln?Thr?Pro?Ile?Lys?Asn?Leu?Tyr?Leu?Thr

500?????????????????505?????????????????510

Gly?Ala?Gly?Thr?His?Pro?Gly?Gly?Ser?Ile?Ser?Gly?Met?Pro?Gly?Arg

515?????????????????520?????????????????525

Asn?Cys?Ala?Arg?Val?Phe?Leu?Lys?Gln?Gln?Arg?Arg?Phe?Trp

530?????????????????535?????????????????540

<210>13

<211>776

<212>DNA

＜213〉living slowly rhizobium (Bradyrhizobium sp.)

<220>

<221>CDS

<222>(1)..(774)

<223>

<400>13

atg?cat?gca?gca?acc?gcc?aag?gct?act?gag?ttc?ggg?gcc?tct?cgg?cgc????????48

Met?His?Ala?Ala?Thr?Ala?Lys?Ala?Thr?Glu?Phe?Gly?Ala?Ser?Arg?Arg

1???????????????5???????????????????10??????????????????15

gac?gat?gcg?agg?cag?cgc?cgc?gtc?ggt?ctc?acg?ctg?gcc?gcg?gtc?atc????????96

Asp?Asp?Ala?Arg?Gln?Arg?Arg?Val?Gly?Leu?Thr?Leu?Ala?Ala?Val?Ile

20??????????????????25??????????????????30

atc?gcc?gcc?tgg?ctg?gtg?ctg?cat?gtc?ggt?ctg?atg?ttc?ttc?tgg?ccg???????144

Ile?Ala?Ala?Trp?Leu?Val?Leu?His?Val?Gly?Leu?Met?Phe?Phe?Trp?Pro

35??????????????????40??????????????????45

ctg?acc?ctt?cac?agc?ctg?ctg?ccg?gct?ttg?cct?ctg?gtg?gtg?ctg?cag???????192

Leu?Thr?Leu?His?Ser?Leu?Leu?Pro?Ala?Leu?Pro?Leu?Val?Val?Leu?Gln

50??????????????????55??????????????????60

acc?tgg?ctc?tat?gta?ggc?ctg?ttc?atc?atc?gcg?cat?gac?tgc?atg?cac???????240

Thr?Trp?Leu?Tyr?Val?Gly?Leu?Phe?Ile?Ile?Ala?His?Asp?Cys?Met?His

65??????????????????70??????????????????75??????????????????80

ggc?tcg?ctg?gtg?ccg?ttc?aag?ccg?cag?gtc?aac?cgc?cgt?atc?gga?cag???????288

Gly?Ser?Leu?Val?Pro?Phe?Lys?Pro?Gln?Val?Asn?Arg?Arg?Ile?Gly?Gln

85??????????????????90??????????????????95

ctc?tgc?ctg?ttc?ctc?tat?gcc?ggg?ttc?tcc?ttc?gac?gct?ctc?aat?gtc???????336

Leu?Cys?Leu?Phe?Leu?Tyr?Ala?Gly?Phe?Ser?Phe?Asp?Ala?Leu?Asn?Val

100?????????????????105?????????????????110

gag?cac?cac?aag?cat?cac?cgc?cat?ccc?ggc?acg?gcc?gag?gat?ccc?gat???????384

Glu?His?His?Lys?His?His?Arg?His?Pro?Gly?Thr?Ala?Glu?Asp?Pro?Asp

115?????????????????120?????????????????125

ttc?gac?gag?gtg?ccg?ccg?cac?ggc?ttc?tgg?cac?tgg?ttc?gcc?agc?ttt???????432

Phe?Asp?Glu?Val?Pro?Pro?His?Gly?Phe?Trp?His?Trp?Phe?Ala?Ser?Phe

130?????????????????135?????????????????140

ttc?ctg?cac?tat?ttc?ggc?tgg?aag?cag?gtc?gcg?atc?atc?gca?gcc?gtc???????480

Phe?Leu?His?Tyr?Phe?Gly?Trp?Lys?Gln?Val?Ala?Ile?Ile?Ala?Ala?Val

145?????????????????150?????????????????155?????????????????160

tcg?ctg?gtt?tat?cag?ctc?gtc?ttc?gcc?gtt?ccc?ttg?cag?aac?atc?ctg???????528

Ser?Leu?Val?Tyr?Gln?Leu?Val?Phe?Ala?Val?Pro?Leu?Gln?Asn?Ile?Leu

165?????????????????170?????????????????175

ctg?ttc?tgg?gcg?ctg?ccc?ggg?ctg?ctg?tcg?gcg?ctg?cag?ctg?ttc?acc???????576

Leu?Phe?Trp?Ala?Leu?Pro?Gly?Leu?Leu?Ser?Ala?Leu?Gln?Leu?Phe?Thr

180?????????????????185?????????????????190

ttc?ggc?acc?tat?ctg?ccg?cac?aag?ccg?gcc?acg?cag?ccc?ttc?gcc?gat???????624

Phe?Gly?Thr?Tyr?Leu?Pro?His?Lys?Pro?Ala?Thr?Gln?Pro?Phe?Ala?Asp

195?????????????????200?????????????????205

cgc?cac?aac?gcg?cgg?acg?agc?gaa?ttt?ccc?gcg?tgg?ctg?tcg?ctg?ctg???????672

Arg?His?Asn?Ala?Arg?Thr?Ser?Glu?Phe?Pro?Ala?Trp?Leu?Ser?Leu?Leu

210?????????????????215?????????????????220

acc?tgc?ttc?cac?ttc?ggc?ttt?cat?cac?gag?cat?cat?ctg?cat?ccc?gat???????720

Thr?Cys?Phe?His?Phe?Gly?Phe?His?His?Glu?His?His?Leu?His?Pro?Asp

225?????????????????230?????????????????235?????????????????240

gcg?ccg?tgg?tgg?cgg?ctg?ccg?gag?atc?aag?cgg?cgg?gcc?ctg?gaa?agg???????768

Ala?Pro?Trp?Trp?Arg?Leu?Pro?Glu?Ile?Lys?Arg?Arg?Ala?Leu?Glu?Arg

245?????????????????250?????????????????255

cgt?gac?ta????????????????????????????????????????????????????????????776

Arg?Asp

<210>14

<211>258

<212>PRT

＜213〉living slowly rhizobium

<400>14

Met?His?Ala?Ala?Thr?Ala?Lys?Ala?Thr?Glu?Phe?Gly?Ala?Ser?Arg?Arg

1???????????????5???????????????????10??????????????????15

Asp?Asp?Ala?Arg?Gln?Arg?Arg?Val?Gly?Leu?Thr?Leu?Ala?Ala?Val?Ile

20??????????????????25??????????????????30

Ile?Ala?Ala?Trp?Leu?Val?Leu?His?Val?Gly?Leu?Met?Phe?Phe?Trp?Pro

35??????????????????40??????????????????????45

Leu?Thr?Leu?His?Ser?Leu?Leu?Pro?Ala?Leu?Pro?Leu?Val?Val?Leu?Gln

50??????????????????55??????????????????60

Thr?Trp?Leu?Tyr?Val?Gly?Leu?Phe?Ile?Ile?Ala?His?Asp?Cys?Met?His

65??????????????????70??????????????????75??????????????????80

Gly?Ser?Leu?Val?Pro?Phe?Lys?Pro?Gln?Val?Asn?Arg?Arg?Ile?Gly?Gln

85??????????????????90??????????????????95

Leu?Cys?Leu?Phe?Leu?Tyr?Ala?Gly?Phe?Ser?Phe?Asp?Ala?Leu?Asn?Val

100?????????????????105?????????????????110

Glu?His?His?Lys?His?His?Arg?His?Pro?Gly?Thr?Ala?Glu?Asp?Pro?Asp

115?????????????????120?????????????????125

Phe?Asp?Glu?Val?Pro?Pro?His?Gly?Phe?Trp?His?Trp?Phe?Ala?Ser?Phe

130?????????????????135?????????????????140

Phe?Leu?His?Tyr?Phe?Gly?Trp?Lys?Gln?Val?Ala?Ile?Ile?Ala?Ala?Val

145?????????????????150?????????????????155?????????????????160

Ser?Leu?Val?Tyr?Gln?Leu?Val?Phe?Ala?Val?Pro?Leu?Gln?Asn?Ile?Leu

165?????????????????170?????????????????175

Leu?Phe?Trp?Ala?Leu?Pro?Gly?Leu?Leu?Ser?Ala?Leu?Gln?Leu?Phe?Thr

180?????????????????185?????????????????190

Phe?Gly?Thr?Tyr?Leu?Pro?His?Lys?Pro?Ala?Thr?Gln?Pro?Phe?Ala?Asp

195?????????????????200?????????????????205

Arg?His?Asn?Ala?Arg?Thr?Ser?Glu?Phe?Pro?Ala?Trp?Leu?Ser?Leu?Leu

210?????????????????215?????????????????220

Thr?Cys?Phe?His?Phe?Gly?Phe?His?His?Glu?His?His?Leu?His?Pro?Asp

225?????????????????230?????????????????235?????????????????240

Ala?Pro?Trp?Trp?Arg?Leu?Pro?Glu?Ile?Lys?Arg?Arg?Ala?Leu?Glu?Arg

245?????????????????250?????????????????255

Arg?Asp

<210>15

<211>777

<212>DNA

＜213〉beads algae (Nostoc sp.)

<220>

<221>CDS

<222>(1)..(777)

<223>

<400>15

atg?gtt?cag?tgt?caa?cca?tca?tct?ctg?cat?tca?gaa?aaa?ctg?gtg?tta????????48

Met?Val?Gln?Cys?Gln?Pro?Ser?Ser?Leu?His?Ser?Glu?Lys?Leu?Val?Leu

1???????????????5???????????????????10??????????????????15

ttg?tca?tcg?aca?atc?aga?gat?gat?aaa?aat?att?aat?aag?ggt?ata?ttt????????96

Leu?Ser?Ser?Thr?Ile?Arg?Asp?Asp?Lys?Asn?Ile?Asn?Lys?Gly?Ile?Phe

20??????????????????25??????????????????30

att?gcc?tgc?ttt?atc?tta?ttt?tta?tgg?gca?att?agt?tta?atc?tta?tta???????144

Ile?Ala?Cys?Phe?Ile?Leu?Phe?Leu?Trp?Ala?Ile?Ser?Leu?Ile?Leu?Leu

35??????????????????40??????????????????45

ctc?tca?ata?gat?aca?tcc?ata?att?cat?aag?agc?tta?tta?ggt?ata?gcc???????192

Leu?Ser?Ile?Asp?Thr?Ser?Ile?Ile?His?Lys?Ser?Leu?Leu?Gly?Ile?Ala

50??????????????????55??????????????????60

atg?ctt?tgg?cag?acc?ttc?tta?tat?aca?ggt?tta?ttt?att?act?gct?cat???????240

Met?Leu?Trp?Gln?Thr?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Ala?His

65??????????????????70??????????????????75??????????????????80

gat?gcc?atg?cac?ggc?gta?gtt?tat?ccc?aaa?aat?ccc?aga?ata?aat?aat???????288

Asp?Ala?Met?His?Gly?Val?Val?Tyr?Pro?Lys?Asn?Pro?Arg?Ile?Asn?Asn

85??????????????????90??????????????????95

ttt?ata?ggt?aag?ctc?act?cta?atc?ttg?tat?gga?cta?ctc?cct?tat?aaa???????336

Phe?Ile?Gly?Lys?Leu?Thr?Leu?Ile?Leu?Tyr?Gly?Leu?Leu?Pro?Tyr?Lys

100?????????????????105?????????????????110

gat?tta?ttg?aaa?aaa?cat?tgg?tta?cac?cac?gga?cat?cct?ggt?act?gat???????384

Asp?Leu?Leu?Lys?Lys?His?Trp?Leu?His?His?Gly?His?Pro?Gly?Thr?Asp

115?????????????????120?????????????????125

tta?gac?cct?gat?tat?tac?aat?ggt?cat?ccc?caa?aac?ttc?ttt?ctt?tgg???????432

Leu?Asp?Pro?Asp?Tyr?Tyr?Asn?Gly?His?Pro?Gln?Asn?Phe?Phe?Leu?Trp

130?????????????????135?????????????????140

tat?cta?cat?ttt?atg?aag?tct?tat?tgg?cga?tgg?acg?caa?att?ttc?gga???????480

Tyr?Leu?His?Phe?Met?Lys?Ser?Tyr?Trp?Arg?Trp?Thr?Gln?Ile?Phe?Gly

145?????????????????150?????????????????155?????????????????160

tta?gtg?atg?att?ttt?cat?gga?ctt?aaa?aat?ctg?gtg?cat?ata?cca?gaa???????528

Leu?Val?Met?Ile?Phe?His?Gly?Leu?Lys?Asn?Leu?Val?His?Ile?Pro?Glu

165?????????????????170?????????????????175

aat?aat?tta?att?ata?ttt?tgg?atg?ata?cct?tct?att?tta?agt?tca?gta???????576

Asn?Asn?Leu?Ile?Ile?Phe?Trp?Met?Ile?Pro?Ser?Ile?Leu?Ser?Ser?Val

180?????????????????185?????????????????190

caa?cta?ttt?tat?ttt?ggt?aca?ttt?ttg?cct?cat?aaa?aag?cta?gaa?ggt???????624

Gln?Leu?Phe?Tyr?Phe?Gly?Thr?Phe?Leu?Pro?His?Lys?Lys?Leu?Glu?Gly

195?????????????????200?????????????????205

ggt?tat?act?aac?ccc?cat?tgt?gcg?cgc?agt?atc?cca?tta?cct?ctt?ttt???????672

Gly?Tyr?Thr?Asn?Pro?His?Cys?Ala?Arg?Ser?Ile?Pro?Leu?Pro?Leu?Phe

210?????????????????215?????????????????220

tgg?tct?ttt?gtt?act?tgt?tat?cac?ttc?ggc?tac?cac?aag?gaa?cat?cac???????720

Trp?Ser?Phe?Val?Thr?Cys?Tyr?His?Phe?Gly?Tyr?His?Lys?Glu?His?His

225?????????????????230?????????????????235?????????????????240

gaa?tac?cct?caa?ctt?cct?tgg?tgg?aaa?tta?cct?gaa?gct?cac?aaa?ata???????768

Glu?Tyr?Pro?Gln?Leu?Pro?Trp?Trp?Lys?Leu?Pro?Glu?Ala?His?Lys?Ile

245?????????????????250?????????????????255

tct?tta?taa???????????????????????????????????????????????????????????777

Ser?Leu

<210>16

<211>258

<212>PRT

＜213〉beads algae

<400>16

Met?Val?Gln?Cys?Gln?Pro?Ser?Ser?Leu?His?Ser?Glu?Lys?Leu?Val?Leu

1???????????????5???????????????????10??????????????????15

Leu?Ser?Ser?Thr?Ile?Arg?Asp?Asp?Lys?Asn?Ile?Asn?Lys?Gly?Ile?Phe

20??????????????????25??????????????????30

Ile?Ala?Cys?Phe?Ile?Leu?Phe?Leu?Trp?Ala?Ile?Ser?Leu?Ile?Leu?Leu

35??????????????????40??????????????????45

Leu?Ser?Ile?Asp?Thr?Ser?Ile?Ile?His?Lys?Ser?Leu?Leu?Gly?Ile?Ala

50??????????????????55??????????????????60

Met?Leu?Trp?Gln?Thr?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Ala?His

65??????????????????70???????????????????75?????????????????80

Asp?Ala?Met?His?Gly?Val?Val?Tyr?Pro?Lys?Asn?Pro?Arg?Ile?Asn?Asn

85??????????????????90??????????????????95

Phe?Ile?Gly?Lys?Leu?Thr?Leu?Ile?Leu?Tyr?Gly?Leu?Leu?Pro?Tyr?Lys

100?????????????????105?????????????????110

Asp?Leu?Leu?Lys?Lys?His?Trp?Leu?His?His?Gly?His?Pro?Gly?Thr?Asp

115?????????????????120?????????????????125

Leu?Asp?Pro?Asp?Tyr?Tyr?Asn?Gly?His?Pro?Gln?Asn?Phe?Phe?Leu?Trp

130?????????????????135?????????????????140

Tyr?Leu?His?Phe?Met?Lys?Ser?Tyr?Trp?Arg?Trp?Thr?Gln?Ile?Phe?Gly

145?????????????????150?????????????????155?????????????????160

Leu?Val?Met?Ile?Phe?His?Gly?Leu?Lys?Asn?Leu?Val?His?Ile?Pro?Glu

165?????????????????170?????????????????175

Asn?Asn?Leu?Ile?Ile?Phe?Trp?Met?Ile?Pro?Ser?Ile?Leu?Ser?Ser?Val

180?????????????????185?????????????????190

Gln?Leu?Phe?Tyr?Phe?Gly?Thr?Phe?Leu?Pro?His?Lys?Lys?Leu?Glu?Gly

195?????????????????200?????????????????205

Gly?Tyr?Thr?Asn?Pro?His?Cys?Ala?Arg?Ser?Ile?Pro?Leu?Pro?Leu?Phe

210?????????????????215?????????????????220

Trp?Ser?Phe?Val?Thr?Cys?Tyr?His?Phe?Gly?Tyr?His?Lys?Glu?His?His

225?????????????????230?????????????????235?????????????????240

Glu?Tyr?Pro?Gln?Leu?Pro?Trp?Trp?Lys?Leu?Pro?Glu?Ala?His?Lys?Ile

245?????????????????250?????????????????255

Ser?Leu

<210>17

<211>1608

<212>DNA

＜213〉haematococcus pluvialis

<220>

<221>CDS

<222>(3)..(971)

<223>

<400>17

ct?aca?ttt?cac?aag?ccc?gtg?agc?ggt?gca?agc?gct?ctg?ccc?cac?atc?????????47

Thr?Phe?His?Lys?Pro?Val?Ser?Gly?Ala?Ser?Ala?Leu?Pro?His?Ile

1???????????????5???????????????????10??????????????????15

ggc?cca?cct?cct?cat?ctc?cat?cgg?tca?ttt?gct?gct?acc?acg?atg?ctg????????95

Gly?Pro?Pro?Pro?His?Leu?His?Arg?Ser?Phe?Ala?Ala?Thr?Thr?Met?Leu

20??????????????????25??????????????????30

tcg?aag?ctg?cag?tca?atc?agc?gtc?aag?gcc?cgc?cgc?gtt?gaa?cta?gcc???????143

Ser?Lys?Leu?Gln?Ser?Ile?Ser?Val?Lys?Ala?Arg?Arg?Val?Glu?Leu?Ala

35??????????????????40??????????????????45

cgc?gac?atc?acg?cgg?ccc?aaa?gtc?tgc?ctg?cat?gct?cag?cgg?tgc?tcg???????191

Arg?Asp?Ile?Thr?Arg?Pro?Lys?Val?Cys?Leu?His?Ala?Gln?Arg?Cys?Ser

50??????????????????55??????????????????60

tta?gtt?cgg?ctg?cga?gtg?gca?gca?cca?cag?aca?gag?gag?gcg?ctg?gga???????239

Leu?Val?Arg?Leu?Arg?Val?Ala?Ala?Pro?Gln?Thr?Glu?Glu?Ala?Leu?Gly

65??????????????????70??????????????????75

acc?gtg?cag?gct?gcc?ggc?gcg?ggc?gat?gag?cac?agc?gcc?gat?gta?gca???????287

Thr?Val?Gln?Ala?Ala?Gly?Ala?Gly?Asp?Glu?His?Ser?Ala?Asp?Val?Ala

80??????????????????85??????????????????90??????????????????95

ctc?cag?cag?ctt?gac?cgg?gct?atc?gca?gag?cgt?cgt?gcc?cgg?cgc?aaa???????335

Leu?Gln?Gln?Leu?Asp?Arg?Ala?Ile?Ala?Glu?Arg?Arg?Ala?Arg?Arg?Lys

100?????????????????105?????????????????110

cgg?gag?cag?ctg?tca?tac?cag?gct?gcc?gcc?att?gca?gca?tca?att?ggc???????383

Arg?Glu?Gln?Leu?Ser?Tyr?Gln?Ala?Ala?Ala?Ile?Ala?Ala?Ser?Ile?Gly

115?????????????????120?????????????????125

gtg?tca?ggc?att?gcc?atc?ttc?gcc?acc?tac?ctg?aga?ttt?gcc?atg?cac???????431

Val?Ser?Gly?Ile?Ala?Ile?Phe?Ala?Thr?Tyr?Leu?Arg?Phe?Ala?Met?His

130?????????????????135?????????????????140

atg?acc?gtg?ggc?ggc?gca?gtg?cca?tgg?ggt?gaa?gtg?gct?ggc?act?ctc???????479

Met?Thr?Val?Gly?Gly?Ala?Val?Pro?Trp?Gly?Glu?Val?Ala?Gly?Thr?Leu

145?????????????????150?????????????????155

ctc?ttg?gtg?gtt?ggt?ggc?gcg?ctc?ggc?atg?gag?atg?tat?gcc?cgc?tat???????527

Leu?Leu?Val?Val?Gly?Gly?Ala?Leu?Gly?Met?Glu?Met?Tyr?Ala?Arg?Tyr

160?????????????????165?????????????????170?????????????????175

gca?cac?aaa?gcc?atc?tgg?cat?gag?tcg?cct?ctg?ggc?tgg?ctg?ctg?cac???????575

Ala?His?Lys?Ala?Ile?Trp?His?Glu?Ser?Pro?Leu?Gly?Trp?Leu?Leu?His

180?????????????????185?????????????????190

aag?agc?cac?cac?aca?cct?cgc?act?gga?ccc?ttt?gaa?gcc?aac?gac?ttg???????623

Lys?Ser?His?His?Thr?Pro?Arg?Thr?Gly?Pro?Phe?Glu?Ala?Asn?Asp?Leu

195?????????????????200?????????????????205

ttt?gca?atc?atc?aat?gga?ctg?ccc?gcc?atg?ctc?ctg?tgt?acc?ttt?ggc???????671

Phe?Ala?Ile?Ile?Asn?Gly?Leu?Pro?Ala?Met?Leu?Leu?Cys?Thr?Phe?Gly

210?????????????????215?????????????????220

ttc?tgg?ctg?ccc?aac?gtc?ctg?ggg?gcg?gcc?tgc?ttt?gga?gcg?ggg?ctg???????719

Phe?Trp?Leu?Pro?Asn?Val?Leu?Gly?Ala?Ala?Cys?Phe?Gly?Ala?Gly?Leu

225?????????????????230?????????????????235

ggc?arc?acg?cta?tac?ggc?atg?gca?tat?atg?ttt?gta?cac?gat?ggc?ctg???????767

Gly?Ile?Thr?Leu?Tyr?Gly?Met?Ala?Tyr?Met?Phe?Val?His?Asp?Gly?Leu

240?????????????????245?????????????????250?????????????????255

gtg?cac?agg?cgc?ttt?ccc?acc?ggg?ccc?atc?gct?ggc?ctg?ccc?tac?atg???????815

Val?His?Arg?Arg?Phe?Pro?Thr?Gly?Pro?Ile?Ala?Gly?Leu?Pro?Tyr?Met

260?????????????????265?????????????????270

aag?cgc?ctg?aca?gtg?gcc?cac?cag?cta?cac?cac?agc?ggc?aag?tac?ggt???????863

Lys?Arg?Leu?Thr?Val?Ala?His?Gln?Leu?His?His?Ser?Gly?Lys?Tyr?Gly

275?????????????????280?????????????????285

ggc?gcg?ccc?tgg?ggt?atg?ttc?ttg?ggt?cca?cag?gag?ctg?cag?cac?att???????911

Gly?Ala?Pro?Trp?Gly?Met?Phe?Leu?Gly?Pro?Gln?Glu?Leu?Gln?His?Ile

290?????????????????295?????????????????300

cca?ggt?gcg?gcg?gag?gag?gtg?gag?cga?ctg?gtc?ctg?gaa?ctg?gac?tgg???????959

Pro?Gly?Ala?Ala?Glu?Glu?Val?Glu?Arg?Leu?Val?Leu?Glu?Leu?Asp?Trp

305?????????????????310?????????????????315

tcc?aag?cgg?tag?ggtgcggaac?caggcacgct?ggtttcacac?ctcatgcctg??????????1011

Ser?Lys?Arg

320

tgataaggtg?tggctagagc?gatgcgtgtg?agacgggtat?gtcacggtcg?actggtctga????1071

tggccaatgg?catcggccat?gtctggtcat?cacgggctgg?ttgcctgggt?gaaggtgatg????1131

cacatcatca?tgtgcggttg?gaggggctgg?cacagtgtgg?gctgaactgg?agcagttgtc????1191

caggctggcg?ttgaatcagt?gagggtttgt?gattggcggt?tgtgaagcaa?tgactccgcc????1251

catattctat?ttgtgggagc?tgagatgatg?gcatgcttgg?gatgtgcatg?gatcatggta????1311

gtgcagcaaa?ctatattcac?ctagggctgt?tggtaggatc?aggtgaggcc?ttgcacattg????1371

catgatgtac?tcgtcatggt?gtgttggtga?gaggatggat?gtggatggat?gtgtattctc????1431

agacgtagac?cttgactgga?ggcttgatcg?agagagtggg?ccgtattctt?tgagagggga????1491

ggctcgtgcc?agaaatggtg?agtggatgac?tgtgacgctg?tacattgcag?gcaggtgaga????1551

tgcactgtct?cgattgtaaa?atacattcag?atgcaaaaaa?aaaaaaaaaa?aaaaaaa???????1608

<210>18

<211>322

<212>PRT

＜213〉haematococcus pluvialis

<400>18

Thr?Phe?His?Lys?Pro?Val?Ser?Gly?Ala?Ser?Ala?Leu?Pro?His?Ile?Gly

1???????????????5???????????????????10??????????????????15

Pro?Pro?Pro?His?Leu?His?Arg?Ser?Phe?Ala?Ala?Thr?Thr?Met?Leu?Ser

20??????????????????25??????????????????30

Lys?Leu?Gln?Ser?Ile?Ser?Val?Lys?Ala?Arg?Arg?Val?Glu?Leu?Ala?Arg

35??????????????????40??????????????????45

Asp?Ile?Thr?Arg?Pro?Lys?Val?Cys?Leu?His?Ala?Gln?Arg?Cys?Ser?Leu

50??????????????????55??????????????????60

Val?Arg?Leu?Arg?Val?Ala?Ala?Pro?Gln?Thr?Glu?Glu?Ala?Leu?Gly?Thr

65??????????????????70??????????????????75??????????????80

Val?Gln?Ala?Ala?Gly?Ala?Gly?Asp?Glu?His?Ser?Ala?Asp?Val?Ala?Leu

85??????????????????90??????????????????95

Gln?Gln?Leu?Asp?Arg?Ala?Ile?Ala?Glu?Arg?Arg?Ala?Arg?Arg?Lys?Arg

100?????????????????105?????????????????110

Glu?Gln?Leu?Ser?Tyr?Gln?Ala?Ala?Ala?Ile?Ala?Ala?Ser?Ile?Gly?Val

115?????????????????120?????????????????125

Ser?Gly?Ile?Ala?Ile?Phe?Ala?Thr?Tyr?Leu?Arg?Phe?Ala?Met?His?Met

130?????????????????135?????????????????140

Thr?Val?Gly?Gly?Ala?Val?Pro?Trp?Gly?Glu?Val?Ala?Gly?Thr?Leu?Leu

145?????????????????150?????????????????155?????????????????160

Leu?Val?Val?Gly?Gly?Ala?Leu?Gly?Met?Glu?Met?Tyr?Ala?Arg?Tyr?Ala

165?????????????????170?????????????????175

His?Lys?Ala?Ile?Trp?His?Glu?Ser?Pro?Leu?Gly?Trp?Leu?Leu?His?Lys

180?????????????????185?????????????????190

Ser?His?His?Thr?Pro?Arg?Thr?Gly?Pro?Phe?Glu?Ala?Asn?Asp?Leu?Phe

195?????????????????200?????????????????205

Ala?Ile?Ile?Asn?Gly?Leu?Pro?Ala?Met?Leu?Leu?Cys?Thr?Phe?Gly?Phe

210?????????????????215?????????????????220

Trp?Leu?Pro?Asn?Val?Leu?Gly?Ala?Ala?Cys?Phe?Gly?Ala?Gly?Leu?Gly

225?????????????????230?????????????????235?????????????????240

Ile?Thr?Leu?Tyr?Gly?Met?Ala?Tyr?Met?Phe?Val?His?Asp?Gly?Leu?Val

245?????????????????250?????????????????255

His?Arg?Arg?Phe?Pro?Thr?Gly?Pro?Ile?Ala?Gly?Leu?Pro?Tyr?Met?Lys

260?????????????????265?????????????????270

Arg?Leu?Thr?Val?Ala?His?Gln?Leu?His?His?Ser?Gly?Lys?Tyr?Gly?Gly

275?????????????????280?????????????????285

Ala?Pro?Trp?Gly?Met?Phe?Leu?Gly?Pro?Gln?Glu?Leu?Gln?His?Ile?Pro

290?????????????????295?????????????????300

Gly?Ala?Ala?Glu?Glu?Val?Glu?Arg?Leu?Val?Leu?Glu?Leu?Asp?Trp?Ser

305?????????????????310?????????????????315?????????????????320

Lys?Arg

<210>19

<211>1503

<212>DNA

＜213〉tomato

<220>

<221>CDS

<222>(1)..(1503)

<223>

<400>19

atg?gat?act?ttg?ttg?aaa?acc?cca?aat?aac?ctt?gaa?ttt?ctg?aac?cca????????48

Met?Asp?Thr?Leu?Leu?Lys?Thr?Pro?Asn?Asn?Leu?Glu?Phe?Leu?Asn?Pro

1???????????????5???????????????????10??????????????????15

cat?cat?ggt?ttt?gct?gtt?aaa?gct?agt?acc?ttt?aga?tct?gag?aag?cat????????96

His?His?Gly?Phe?Ala?Val?Lys?Ala?Ser?Thr?Phe?Arg?Ser?Glu?Lys?His

20??????????????????25??????????????????30

cat?aat?ttt?ggt?tct?agg?aag?ttt?tgt?gaa?act?ttg?ggt?aga?agt?gtt???????144

His?Asn?Phe?Gly?Ser?Arg?Lys?Phe?Cys?Glu?Thr?Leu?Gly?Arg?Ser?Val

35??????????????????40??????????????????45

tgt?gtt?aag?ggt?agt?agt?agt?gct?ctt?tta?gag?ctt?gta?cct?gag?acc???????192

Cys?Val?Lys?Gly?Ser?Ser?Ser?Ala?Leu?Leu?Glu?Leu?Val?Pro?Glu?Thr

50??????????????????55??????????????????60

aaa?aag?gag?aat?ctt?gat?ttt?gag?ctt?cct?atg?tat?gac?cct?tca?aaa???????240

Lys?Lys?Glu?Asn?Leu?Asp?Phe?Glu?Leu?Pro?Met?Tyr?Asp?Pro?Ser?Lys

65??????????????????70??????????????????75??????????????????80

ggg?gtt?gtt?gtg?gat?ctt?gct?gtg?gtt?ggt?ggt?ggc?cct?gca?gga?ctt???????288

Gly?Val?Val?Val?Asp?Leu?Ala?Val?Val?Gly?Gly?Gly?Pro?Ala?Gly?Leu

85??????????????????90??????????????????95

gct?gtt?gca?cag?caa?gtt?tct?gaa?gca?gga?ctc?tct?gtt?tgt?tca?att???????336

Ala?Val?Ala?Gln?Gln?Val?Ser?Glu?Ala?Gly?Leu?Ser?Val?Cys?Ser?Ile

100?????????????????105?????????????????110

gat?ccg?aat?cct?aaa?ttg?ata?tgg?cct?aat?aac?tat?ggt?gtt?tgg?gtg???????384

Asp?Pro?Asn?Pro?Lys?Leu?Ile?Trp?Pro?Asn?Asn?Tyr?Gly?Val?Trp?Val

115?????????????????120?????????????????125

gat?gaa?ttt?gag?gct?atg?gac?ttg?tta?gat?tgt?cta?gat?gct?acc?tgg???????432

Asp?Glu?Phe?Glu?Ala?Met?Asp?Leu?Leu?Asp?Cys?Leu?Asp?Ala?Thr?Trp

130?????????????????135?????????????????140

tct?ggt?gca?gca?gtg?tac?att?gat?gat?aat?acg?gct?aaa?gat?ctt?cat???????480

Ser?Gly?Ala?Ala?Val?Tyr?Ile?Asp?Asp?Asn?Thr?Ala?Lys?Asp?Leu?His

145?????????????????150?????????????????155?????????????????160

aga?cct?tat?gga?agg?gtt?aac?cgg?aaa?cag?ctg?aaa?tcg?aaa?atg?atg???????528

Arg?Pro?Tyr?Gly?Arg?Val?Asn?Arg?Lys?Gln?Leu?Lys?Ser?Lys?Met?Met

165?????????????????170?????????????????175

cag?aaa?tgt?ata?atg?aat?ggt?gtt?aaa?ttc?cac?caa?gcc?aaa?gtt?ata???????576

Gln?Lys?Cys?Ile?Met?Asn?Gly?Val?Lys?Phe?His?Gln?Ala?Lys?Val?Ile

180?????????????????185?????????????????190

aag?gtg?att?cat?gag?gaa?tcg?aaa?tcc?atg?ttg?ata?tgc?aat?gat?ggt???????624

Lys?Val?Ile?His?Glu?Glu?Ser?Lys?Ser?Met?Leu?Ile?Cys?Asn?Asp?Gly

195?????????????????200?????????????????205

att?act?att?cag?gca?acg?gtg?gtg?ctc?gat?gca?act?ggc?ttc?tct?aga???????672

Ile?Thr?Ile?Gln?Ala?Thr?Val?Val?Leu?Asp?Ala?Thr?Gly?Phe?Ser?Arg

210?????????????????215?????????????????220

tct?ctt?gtt?cag?tat?gat?aag?cct?tat?aac?ccc?ggg?tat?caa?gtt?gct???????720

Ser?Leu?Val?Gln?Tyr?Asp?Lys?Pro?Tyr?Asn?Pro?Gly?Tyr?Gln?Val?Ala

225?????????????????230?????????????????235?????????????????240

tat?ggc?att?ttg?gct?gaa?gtg?gaa?gag?cac?ccc?ttt?gat?gta?aac?aag???????768

Tyr?Gly?Ile?Leu?Ala?Glu?Val?Glu?Glu?His?Pro?Phe?Asp?Val?Asn?Lys

245?????????????????250?????????????????255

atg?gtt?ttc?atg?gat?tgg?cga?gat?tct?cat?ttg?aag?aac?aat?act?gat???????816

Met?Val?Phe?Met?Asp?Trp?Arg?Asp?Ser?His?Leu?Lys?Asn?Asn?Thr?Asp

260?????????????????265?????????????????270

ctc?aag?gag?aga?aat?agt?aga?ata?cca?act?ttt?ctt?tat?gca?atg?cca???????864

Leu?Lys?Glu?Arg?Asn?Ser?Arg?Ile?Pro?Thr?Phe?Leu?Tyr?Ala?Met?Pro

275?????????????????280?????????????????285

ttt?tca?tcc?aac?agg?ata?ttt?ctt?gaa?gaa?aca?tca?ctc?gta?gct?cgt???????912

Phe?Ser?Ser?Asn?Arg?Ile?Phe?Leu?Glu?Glu?Thr?Ser?Leu?Val?Ala?Arg

290????????????????295?????????????????300

cct?ggc?ttg?cgt?ata?gat?gat?att?caa?gaa?cga?atg?gtg?gct?cgt?tta???????960

Pro?Gly?Leu?Arg?Ile?Asp?Asp?Ile?Gln?Glu?Arg?Met?Val?Ala?Arg?Leu

305?????????????????310?????????????????315?????????????????320

aac?cat?ttg?ggg?ata?aaa?gtg?aag?agc?att?gaa?gaa?gat?gaa?cat?tgt??????1008

Asn?His?Leu?Gly?Ile?Lys?Val?Lys?Ser?Ile?Glu?Glu?Asp?Glu?His?Cys

325?????????????????330?????????????????335

cta?ata?cca?atg?ggt?ggt?cca?ctt?cca?gta?tta?cct?cag?aga?gtc?gtt??????1056

Leu?Ile?Pro?Met?Gly?Gly?Pro?Leu?Pro?Val?Leu?Pro?Gln?Arg?Val?Val

340?????????????????345?????????????????350

gga?atc?ggt?ggt?aca?gct?ggc?atg?gtt?cat?cca?tcc?acc?ggt?tat?atg??????1104

Gly?Ile?Gly?Gly?Thr?Ala?Gly?Met?Val?His?Pro?Ser?Thr?Gly?Tyr?Met

355?????????????????360?????????????????365

gtg?gca?agg?aca?cta?gct?gcg?gct?cct?gtt?gtt?gcc?aat?gcc?ata?att??????1152

Val?Ala?Arg?Thr?Leu?Ala?Ala?Ala?Pro?Val?Val?Ala?Asn?Ala?Ile?Ile

370?????????????????375?????????????????380

caa?tac?ctc?ggt?tct?gaa?aga?agt?cat?tcg?ggt?aat?gaa?tta?tcc?aca??????1200

Gln?Tyr?Leu?Gly?Ser?Glu?Arg?Ser?His?Ser?Gly?Asn?Glu?Leu?Ser?Thr

385?????????????????390?????????????????395?????????????????400

gct?gtt?tgg?aaa?gat?ttg?tgg?cct?ata?gag?agg?aga?cgt?caa?aga?gag??????1248

Ala?Val?Trp?Lys?Asp?Leu?Trp?Pro?Ile?Glu?Arg?Arg?Arg?Gln?Arg?Glu

405?????????????????410?????????????????415

ttc?ttc?tgc?ttc?ggt?atg?gat?att?ctt?ctg?aag?ctt?gat?tta?cct?gct??????1296

Phe?Phe?Cys?Phe?Gly?Met?Asp?Ile?Leu?Leu?Lys?Leu?Asp?Leu?Pro?Ala

420?????????????????425?????????????????430

aca?aga?agg?ttc?ttt?gat?gca?ttc?ttt?gac?tta?gaa?cct?cgt?tat?tgg??????1344

Thr?Arg?Arg?Phe?Phe?Asp?Ala?Phe?Phe?Asp?Leu?Glu?Pro?Arg?Tyr?Trp

435?????????????????440?????????????????445

cat?ggc?ttc?tta?tcg?tct?cga?ttg?ttt?cta?cct?gaa?ctc?ata?gtt?ttt??????1392

His?Gly?Phe?Leu?Ser?Ser?Arg?Leu?Phe?Leu?Pro?Glu?Leu?Ile?Val?Phe

450?????????????????455?????????????????460

ggg?ctg?tct?cta?ttc?tct?cat?gct?tca?aat?act?tct?aga?ttt?gag?ata??????1440

Gly?Leu?Ser?Leu?Phe?Ser?His?Ala?Ser?Asn?Thr?Ser?Arg?Phe?Glu?Ile

465?????????????????470?????????????????475?????????????????480

atg?aca?aag?gga?act?gtt?cca?tta?gta?aat?atg?atc?aac?aat?ttg?tta??????1488

Met?Thr?Lys?Gly?Thr?Val?Pro?Leu?Val?Asn?Met?Ile?Asn?Asn?Leu?Leu

485?????????????????490?????????????????495

cag?gat?aaa?gaa?tga??????????????????????????????????????????????????1503

Gln?Asp?Lys?Glu

500

<210>20

<211>500

<212>PRT

＜213〉tomato

<400>20

Met?Asp?Thr?Leu?Leu?Lys?Thr?Pro?Asn?Asn?Leu?Glu?Phe?Leu?Asn?Pro

1???????????????5???????????????????10??????????????????15

His?His?Gly?Phe?Ala?Val?Lys?Ala?Ser?Thr?Phe?Arg?Ser?Glu?Lys?His

20??????????????????25??????????????????30

His?Asn?Phe?Gly?Ser?Arg?Lys?Phe?Cys?Glu?Thr?Leu?Gly?Arg?Ser?Val

35??????????????????40??????????????????45

Cys?Val?Lys?Gly?Ser?Ser?Ser?Ala?Leu?Leu?Glu?Leu?Val?Pro?Glu?Thr

50??????????????????55??????????????????60

Lys?Lys?Glu?Asn?Leu?Asp?Phe?Glu?Leu?Pro?Met?Tyr?Asp?Pro?Ser?Lys

65??????????????????70??????????????????75??????????????????80

Gly?Val?Val?Val?Asp?Leu?Ala?Val?Val?Gly?Gly?Gly?Pro?Ala?Gly?Leu

85??????????????????90??????????????????95

Ala?Val?Ala?Gln?Gln?Val?Ser?Glu?Ala?Gly?Leu?Ser?Val?Cys?Ser?Ile

100?????????????????105?????????????????110

Asp?Pro?Asn?Pro?Lys?Leu?Ile?Trp?Pro?Asn?Asn?Tyr?Gly?Val?Trp?Val

115?????????????????120?????????????????125

Asp?Glu?Phe?Glu?Ala?Met?Asp?Leu?Leu?Asp?Cys?Leu?Asp?Ala?Thr?Trp

130?????????????????135?????????????????140

Ser?Gly?Ala?Ala?Val?Tyr?Ile?Asp?Asp?Asn?Thr?Ala?Lys?Asp?Leu?His

145?????????????????150?????????????????155?????????????????160

Arg?Pro?Tyr?Gly?Arg?Val?Asn?Arg?Lys?Gln?Leu?Lys?Ser?Lys?Met?Met

165?????????????????170?????????????????175

Gln?Lys?Cys?Ile?Met?Asn?Gly?Val?Lys?Phe?His?Gln?Ala?Lys?Val?Ile

180?????????????????185?????????????????190

Lys?Val?Ile?His?Glu?Glu?Ser?Lys?Ser?Met?Leu?Ile?Cys?Asn?Asp?Gly

195?????????????????200?????????????????205

Ile?Thr?Ile?Gln?Ala?Thr?Val?Val?Leu?Asp?Ala?Thr?Gly?Phe?Ser?Arg

210?????????????????215?????????????????220

Ser?Leu?Val?Gln?Tyr?Asp?Lys?Pro?Tyr?Asn?Pro?Gly?Tyr?Gln?Val?Ala

225?????????????????230?????????????????235?????????????????240

Tyr?Gly?Ile?Leu?Ala?Glu?Val?Glu?Glu?His?Pro?Phe?Asp?Val?Asn?Lys

245?????????????????250?????????????????255

Met?Val?Phe?Met?Asp?Trp?Arg?Asp?Ser?His?Leu?Lys?Asn?Asn?Thr?Asp

260?????????????????265?????????????????270

Leu?Lys?Glu?Arg?Asn?Ser?Arg?Ile?Pro?Thr?Phe?Leu?Tyr?Ala?Met?Pro

275?????????????????280?????????????????285

Phe?Ser?Ser?Asn?Arg?Ile?Phe?Leu?Glu?Glu?Thr?Ser?Leu?Val?Ala?Arg

290?????????????????295?????????????????300

Pro?Gly?Leu?Arg?Ile?Asp?Asp?Ile?Gln?Glu?Arg?Met?Val?Ala?Arg?Leu

305?????????????????310?????????????????315?????????????????320

Asn?His?Leu?Gly?Ile?Lys?Val?Lys?Ser?Ile?Glu?Glu?Asp?Glu?His?Cys

325?????????????????330?????????????????335

Leu?Ile?Pro?Met?Gly?Gly?Pro?Leu?Pro?Val?Leu?Pro?Gln?Arg?Val?Val

340?????????????????345?????????????????350

Gly?Ile?Gly?Gly?Thr?Ala?Gly?Met?Val?His?Pro?Ser?Thr?Gly?Tyr?Met

355?????????????????360?????????????????365

Val?Ala?Arg?Thr?Leu?Ala?Ala?Ala?Pro?Val?Val?Ala?Asn?Ala?Ile?Ile

370?????????????????375?????????????????380

Gln?Tyr?Leu?Gly?Ser?Glu?Arg?Ser?His?Ser?Gly?Asn?Glu?Leu?Ser?Thr

385?????????????????390?????????????????395?????????????????400

Ala?Val?Trp?Lys?Asp?Leu?Trp?Pro?Ile?Glu?Arg?Arg?Arg?Gln?Arg?Glu

405?????????????????410?????????????????415

Phe?Phe?Cys?Phe?Gly?Met?Asp?Ile?Leu?Leu?Lys?Leu?Asp?Leu?Pro?Ala

420?????????????????425?????????????????430

Thr?Arg?Arg?Phe?Phe?Asp?Ala?Phe?Phe?Asp?Leu?Glu?Pro?Arg?Tyr?Trp

435?????????????????440?????????????????445

His?Gly?Phe?Leu?Ser?Ser?Arg?Leu?Phe?Leu?Pro?Glu?Leu?Ile?Val?Phe

450?????????????????455?????????????????460

Gly?Leu?Ser?Leu?Phe?Ser?His?Ala?Ser?Asn?Thr?Ser?Arg?Phe?Glu?Ile

465?????????????????470?????????????????475?????????????????480

Met?Thr?Lys?Gly?Thr?Val?Pro?Leu?Val?Asn?Met?Ile?Asn?Asn?Leu?Leu

485???????????????490???????????????495

Gln?Asp?Lys?Glu

500

<210>21

<211>195

<212>DNA

＜213〉potato

<220>

＜221〉introne

<222>(1)..(195)

<223>

<400>21

tacgtaagtt?tctgcttcta?cctttgatat?atatataata?attatcatta?attagtagta??????60

atataatatt?tcaaatattt?ttttcaaaat?aaaagaatgt?agtatatagc?aattgctttt?????120

ctgtagttta?taagtgtgta?tattttaatt?tataactttt?ctaatatatg?accaaaattt?????180

gttgatgtgc?agctg??????????????????????????????????????????????????????195

<210>22

<211>1155

<212>DNA

＜213〉haematococcus pluvialis

<220>

<221>CDS

<222>(6)..(995)

<223>

<400>22

gaagc?atg?cag?cta?gca?gcg?aca?gta?atg?ttg?gag?cag?ctt?acc?gga?agc??????50

Met?Gln?Leu?Ala?Ala?Thr?Val?Met?Leu?Glu?Gln?Leu?Thr?Gly?Ser

1???????????????5???????????????????10??????????????????15

gct?gag?gca?ctc?aag?gag?aag?gag?aag?gag?gtt?gca?ggc?agc?tct?gac????????98

Ala?Glu?Ala?Leu?Lys?Glu?Lys?Glu?Lys?Glu?Val?Ala?Gly?Ser?Ser?Asp

20??????????????????25??????????????????30

gtg?ttg?cgt?aca?tgg?gcg?acc?cag?tac?tcg?ctt?ccg?tca?gag?gag?tca???????146

Val?Leu?Arg?Thr?Trp?Ala?Thr?Gln?Tyr?Ser?Leu?Pro?Ser?Glu?Glu?Ser

35??????????????????40??????????????????45

gac?gcg?gcc?cgc?ccg?gga?ctg?aag?aat?gcc?tac?aag?cca?cca?cct?tcc???????194

Asp?Ala?Ala?Arg?Pro?Gly?Leu?Lys?Asn?Ala?Tyr?Lys?Pro?Pro?Pro?Ser

50??????????????????55??????????????????60

gac?aca?aag?ggc?atc?aca?atg?gcg?cta?gct?gtc?atc?ggc?tcc?tgg?gcc???????242

Asp?Thr?Lys?Gly?Ile?Thr?Met?Ala?Leu?Ala?Val?Ile?Gly?Ser?Trp?Ala

65??????????????????70??????????????????75

gca?gtg?ttc?ctc?cac?gcc?att?ttt?caa?atc?aag?ctt?ccg?acc?tcc?ttg???????290

Ala?Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile?Lys?Leu?Pro?Thr?Ser?Leu

80??????????????????85??????????????????90??????????????????95

gac?cag?ctg?cac?tgg?ctg?ccc?gtg?tca?gat?gcc?aca?gct?cag?ctg?gtt???????338

Asp?Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Asp?Ala?Thr?Ala?Gln?Leu?Val

100?????????????????105?????????????????110

agc?ggc?agc?agc?agc?ctg?ctg?cac?atc?gtc?gta?gta?ttc?ttt?gtc?ctg???????386

Ser?Gly?Ser?Ser?Ser?Leu?Leu?His?Ile?Val?Val?Val?Phe?Phe?Val?Leu

115?????????????????120?????????????????125

gag?ttc?ctg?tac?aca?ggc?ctt?ttt?atc?acc?acg?cat?gat?gct?atg?cat???????434

Glu?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Thr?His?Asp?Ala?Met?His

130?????????????????135?????????????????140

ggc?acc?atc?gcc?atg?aga?aac?agg?cag?ctt?aat?gac?ttc?ttg?ggc?aga???????482

Gly?Thr?Ile?Ala?Met?Arg?Asn?Arg?Gln?Leu?Asn?Asp?Phe?Leu?Gly?Arg

145?????????????????150?????????????????155

gta?tgc?atc?tcc?ttg?tac?gcc?tgg?ttt?gat?tac?aac?atg?ctg?cac?cgc???????530

Val?Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp?Tyr?Asn?Met?Leu?His?Arg

160?????????????????165?????????????????170?????????????????175

aag?cat?tgg?gag?cac?cac?aac?cac?act?ggc?gag?gtg?ggc?aag?gac?cct???????578

Lys?His?Trp?Glu?His?His?Asn?His?Thr?Gly?Glu?Val?Gly?Lys?Asp?Pro

180?????????????????185?????????????????190

gac?ttc?cac?agg?gga?aac?cct?ggc?att?gtg?ccc?tgg?ttt?gcc?agc?ttc???????626

Asp?Phe?His?Arg?Gly?Asn?Pro?Gly?Ile?Val?Pro?Trp?Phe?Ala?Ser?Phe

195?????????????????200?????????????????205

atg?tcc?agc?tac?atg?tcg?atg?tgg?cag?ttt?gcg?cgc?ctc?gca?tgg?tgg???????674

Met?Ser?Ser?Tyr?Met?Ser?Met?Trp?Gln?Phe?Ala?Arg?Leu?Ala?Trp?Trp

210?????????????????215?????????????????220

acg?gtg?gtc?atg?cag?ctg?ctg?ggt?gcg?cca?atg?gcg?aac?ctg?ctg?gtg???????722

Thr?Val?Val?Met?Gln?Leu?Leu?Gly?Ala?Pro?Met?Ala?Asn?Leu?Leu?Val

225?????????????????230?????????????????235

ttc?atg?gcg?gcc?gcg?ccc?atc?ctg?tcc?gcc?ttc?cgc?ttg?ttc?tac?ttt???????770

Phe?Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu?Phe?Tyr?Phe

240?????????????????245?????????????????250?????????????????255

ggc?acg?tac?atg?ccc?cac?aag?cct?gag?cct?ggc?gcc?gcg?tca?ggc?tct???????818

Gly?Thr?Tyr?Met?Pro?His?Lys?Pro?Glu?Pro?Gly?Ala?Ala?Ser?Gly?Ser

260?????????????????265?????????????????270

tca?cca?gcc?gtc?atg?aac?tgg?tgg?aag?tcg?cgc?act?agc?cag?gcg?tcc???????866

Ser?Pro?Ala?Val?Met?Asn?Trp?Trp?Lys?Ser?Arg?Thr?Ser?Gln?Ala?Ser

275?????????????????280?????????????????285

gac?ctg?gtc?agc?ttt?ctg?acc?tgc?tac?cac?ttc?gac?ctg?cac?tgg?gag???????914

Asp?Leu?Val?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?His?Trp?Glu

290?????????????????295?????????????????300

cac?cac?cgc?tgg?ccc?ttt?gcc?ccc?tgg?tgg?gag?ctg?ccc?aac?tgc?cgc???????962

His?His?Arg?Trp?Pro?Phe?Ala?Pro?Trp?Trp?Glu?Leu?Pro?Asn?Cys?Arg

305?????????????????310?????????????????315

cgc?ctg?tct?ggc?cga?ggt?ctg?gtt?cct?gcc?tag?ctggacacac?tgcagtgggc????1015

Arg?Leu?Ser?Gly?Arg?Gly?Leu?Val?Pro?Ala

320?????????????????325

cctgctgcca?gctgggcatg?caggttgtgg?caggactggg?tgaggtgaaa?agctgcaggc????1075

gctgctgccg?gacacgctgc?atgggctacc?ctgtgtagct?gccgccacta?ggggaggggg????1135

tttgtagctg?tcgagcttgc????????????????????????????????????????????????1155

<210>23

<211>329

<212>PRT

＜213〉haematococcus pluvialis

<400>23

Met?Gln?Leu?Ala?Ala?Thr?Val?Met?Leu?Glu?Gln?Leu?Thr?Gly?Ser?Ala

1???????????????5???????????????????10??????????????????15

Glu?Ala?Leu?Lys?Glu?Lys?Glu?Lys?Glu?Val?Ala?Gly?Ser?Ser?Asp?Val

20??????????????????25??????????????????30

Leu?Arg?Thr?Trp?Ala?Thr?Gln?Tyr?Ser?Leu?Pro?Ser?Glu?Glu?Ser?Asp

35??????????????????40??????????????????45

Ala?Ala?Arg?Pro?Gly?Leu?Lys?Asn?Ala?Tyr?Lys?Pro?Pro?Pro?Ser?Asp

50??????????????????55??????????????????60

Thr?Lys?Gly?Ile?Thr?Met?Ala?Leu?Ala?Val?Ile?Gly?Ser?Trp?Ala?Ala

65??????????????????70??????????????????75??????????????????80

Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile?Lys?Leu?Pro?Thr?Ser?Leu?Asp

85??????????????????90??????????????????95

Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Asp?Ala?Thr?Ala?Gln?Leu?Val?Ser

100?????????????????105?????????????????110

Gly?Ser?Ser?Ser?Leu?Leu?His?Ile?Val?Val?Val?Phe?Phe?Val?Leu?Glu

115?????????????????120?????????????????125

Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Thr?His?Asp?Ala?Met?His?Gly

130?????????????????135?????????????????140

Thr?Ile?Ala?Met?Arg?Asn?Arg?Gln?Leu?Asn?Asp?Phe?Leu?Gly?Arg?Val

145?????????????????150?????????????????155?????????????????160

Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp?Tyr?Asn?Met?Leu?His?Arg?Lys

165?????????????????170?????????????????175

His?Trp?Glu?His?His?Asn?His?Thr?Gly?Glu?Val?Gly?Lys?Asp?Pro?Asp

180?????????????????185?????????????????190

Phe?His?Arg?Gly?Asn?Pro?Gly?Ile?Val?Pro?Trp?Phe?Ala?Ser?Phe?Met

195?????????????????200?????????????????205

Ser?Ser?Tyr?Met?Ser?Met?Trp?Gln?Phe?Ala?Arg?Leu?Ala?Trp?Trp?Thr

210?????????????????215?????????????????220

Val?Val?Met?Gln?Leu?Leu?Gly?Ala?Pro?Met?Ala?Asn?Leu?Leu?Val?Phe

225?????????????????230?????????????????235?????????????????240

Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu?Phe?Tyr?Phe?Gly

245?????????????????250?????????????????255

Thr?Tyr?Met?Pro?His?Lys?Pro?Glu?Pro?Gly?Ala?Ala?Ser?Gly?Ser?Ser

260?????????????????265?????????????????270

Pro?Ala?Val?Met?Asn?Trp?Trp?Lys?Ser?Arg?Thr?Ser?Gln?Ala?Ser?Asp

275?????????????????280?????????????????285

Leu?Val?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?His?Trp?Glu?His

290?????????????????295?????????????????300

His?Arg?Trp?Pro?Phe?Ala?Pro?Trp?Trp?Glu?Leu?Pro?Asn?Cys?Arg?Arg

305?????????????????310?????????????????315?????????????????320

Leu?Ser?Gly?Arg?Gly?Leu?Val?Pro?Ala

325

<210>24

<211>1111

<212>DNA

＜213〉haematococcus pluvialis

<220>

<221>CDS

<222>(4)..(951)

<223>

<400>24

tgc?atg?cta?gag?gca?ctc?aag?gag?aag?gag?aag?gag?gtt?gca?ggc?agc????????48

Met?Leu?Glu?Ala?Leu?Lys?Glu?Lys?Glu?Lys?Glu?Val?Ala?Gly?Ser

1???????????????5???????????????????10??????????????????15

tct?gac?gtg?ttg?cgt?aca?tgg?gcg?acc?cag?tac?tcg?ctt?ccg?tca?gaa????????96

Ser?Asp?Val?Leu?Arg?Thr?Trp?Ala?Thr?Gln?Tyr?Ser?Leu?Pro?Ser?Glu

20??????????????????25??????????????????30

gag?tca?gac?gcg?gcc?cgc?ccg?gga?ctg?aag?aat?gcc?tac?aag?cca?cca???????144

Glu?Ser?Asp?Ala?Ala?Arg?Pro?Gly?Leu?Lys?Asn?Ala?Tyr?Lys?Pro?Pro

35??????????????????40??????????????????45

cct?tcc?gac?aca?aag?ggc?atc?aca?atg?gcg?cta?gct?gtc?atc?ggc?tcc???????192

Pro?Ser?Asp?Thr?Lys?Gly?Ile?Thr?Met?Ala?Leu?Ala?Val?Ile?Gly?Ser

50??????????????????55??????????????????60

tgg?gcc?gca?gtg?ttc?ctc?cac?gcc?att?ttt?caa?atc?aag?ctt?ccg?acc???????240

Trp?Ala?Ala?Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile?Lys?Leu?Pro?Thr

65??????????????????70??????????????????75

tcc?ttg?gac?cag?ctg?cac?tgg?ctg?ccc?gtg?tca?gat?gcc?aca?gct?cag???????288

Ser?Leu?Asp?Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Asp?Ala?Thr?Ala?Gln

80??????????????????85??????????????????90??????????????????95

ctg?gtt?agc?ggc?agc?agc?agc?ctg?ctg?cac?atc?gtc?gta?gta?ttc?ttt???????336

Leu?Val?Ser?Gly?Ser?Ser?Ser?Leu?Leu?His?Ile?Val?Val?Val?Phe?Phe

100?????????????????105?????????????????110

gtc?ctg?gag?ttc?ctg?tac?aca?ggc?ctt?ttt?atc?acc?acg?cat?gat?gct???????384

Val?Leu?Glu?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Thr?His?Asp?Ala

115?????????????????120?????????????????125

atg?cat?ggc?acc?atc?gcc?atg?aga?aac?agg?cag?ctt?aat?gac?ttc?ttg???????432

Met?His?Gly?Thr?Ile?Ala?Met?Arg?Asn?Arg?Gln?Leu?Asn?Asp?Phe?Leu

130?????????????????135?????????????????140

ggc?aga?gta?tgc?atc?tcc?ttg?tac?gcc?tgg?ttt?gat?tac?aac?atg?ctg???????480

Gly?Arg?Val?Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp?Tyr?Asn?Met?Leu

145?????????????????150?????????????????155

cac?cgc?aag?cat?tgg?gag?cac?cac?aac?cac?act?ggc?gag?gtg?ggc?aag???????528

His?Arg?Lys?His?Trp?Glu?His?His?Asn?His?Thr?Gly?Glu?Val?Gly?Lys

160?????????????????165?????????????????170?????????????????175

gac?cct?gac?ttc?cac?agg?gga?aac?cct?ggc?att?gtg?ccc?tgg?ttt?gcc???????576

Asp?Pro?Asp?Phe?His?Arg?Gly?Asn?Pro?Gly?Ile?Val?Pro?Trp?Phe?Ala

180?????????????????185?????????????????190

agc?ttc?atg?tcc?agc?tac?atg?tcg?atg?tgg?cag?ttt?gcg?cgc?ctc?gca???????624

Ser?Phe?Met?Ser?Ser?Tyr?Met?Ser?Met?Trp?Gln?Phe?Ala?Arg?Leu?Ala

195?????????????????200?????????????????205

tgg?tgg?acg?gtg?gtc?atg?cag?ctg?ctg?ggt?gcg?cca?atg?gcg?aac?ctg???????672

Trp?Trp?Thr?Val?Val?Met?Gln?Leu?Leu?Gly?Ala?Pro?Met?Ala?Asn?Leu

210?????????????????215?????????????????220

ctg?gtg?ttc?atg?gcg?gcc?gcg?ccc?atc?ctg?tcc?gcc?ttc?cgc?ttg?ttc???????720

Leu?Val?Phe?Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu?Phe

225?????????????????230?????????????????235

tac?ttt?ggc?acg?tac?atg?ccc?cac?aag?cct?gag?cct?ggc?gcc?gcg?tca???????768

Tyr?Phe?Gly?Thr?Tyr?Met?Pro?His?Lys?Pro?Glu?Pro?Gly?Ala?Ala?Ser

240?????????????????245?????????????????250?????????????????255

ggc?tct?rca?cca?gcc?gtc?atg?aac?tgg?tgg?aag?tcg?cgc?act?agc?cag???????816

Gly?Ser?Ser?Pro?Ala?Val?Met?Asn?Trp?Trp?Lys?Ser?Arg?Thr?Ser?Gln

260?????????????????265?????????????????270

gcg?tcc?gac?ctg?gtc?agc?ttt?ctg?acc?tgc?tac?cac?ttc?gac?ctg?cac???????864

Ala?Ser?Asp?Leu?Val?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?His

275?????????????????280?????????????????285

tgg?gag?cac?cac?cgc?tgg?ccc?ttc?gcc?ccc?tgg?tgg?gag?ctg?ccc?aac???????912

Trp?Glu?His?His?Arg?Trp?Pro?Phe?Ala?Pro?Trp?Trp?Glu?Leu?Pro?Asn

290?????????????????295?????????????????300

tgc?cgc?cgc?ctg?tct?ggc?cga?ggt?ctg?gtt?cct?gcc?tag?ctggacacac????????961

Cys?Arg?Arg?Leu?Ser?Gly?Arg?Gly?Leu?Val?Pro?Ala

305?????????????????310?????????????????315

tgcagtgggc?cctgctgcca?gctgggcatg?caggttgtgg?caggactggg?tgaggtgaaa????1021

agctgcaggc?gctgctgccg?gacacgttgc?atgggctacc?ctgtgtagct?gccgccacta????1081

ggggaggggg?tttgtagctg?tcgagcttgc?????????????????????????????????????1111

<210>25

<211>315

<212>PRT

＜213〉haematococcus pluvialis

<400>25

Met?Leu?Glu?Ala?Leu?Lys?Glu?Lys?Glu?Lys?Glu?Val?Ala?Gly?Ser?Ser

1???????????????5???????????????????10??????????????????15

Asp?Val?Leu?Arg?Thr?Trp?Ala?Thr?Gln?Tyr?Ser?Leu?Pro?Ser?Glu?Glu

20??????????????????25??????????????????30

Ser?Asp?Ala?Ala?Arg?Pro?Gly?Leu?Lys?Asn?Ala?Tyr?Lys?Pro?Pro?Pro

35??????????????????40??????????????????45

Ser?Asp?Thr?Lys?Gly?Ile?Thr?Met?Ala?Leu?Ala?Val?Ile?Gly?Ser?Trp

50??????????????????55??????????????????60

Ala?Ala?Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile?Lys?Leu?Pro?Thr?Ser

65??????????????????70??????????????????75??????????????????80

Leu?Asp?Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Asp?Ala?Thr?Ala?Gln?Leu

85??????????????????90??????????????????95

Val?Ser?Gly?Ser?Ser?Ser?Leu?Leu?His?Ile?Val?Val?Val?Phe?Phe?Val

100?????????????????105?????????????????????110

Leu?Glu?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Thr?His?Asp?Ala?Met

115?????????????????120?????????????????125

His?Gly?Thr?Ile?Ala?Met?Arg?Asn?Arg?Gln?Leu?Asn?Asp?Phe?Leu?Gly

130?????????????????135?????????????????140

Arg?Val?Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp?Tyr?Asn?Met?Leu?His

145?????????????????150?????????????????155?????????????????160

Arg?Lys?His?Trp?Glu?His?His?Asn?His?Thr?Gly?Glu?Val?Gly?Lys?Asp

165?????????????????170?????????????????175

Pro?Asp?Phe?His?Arg?Gly?Asn?Pro?Gly?Ile?Val?Pro?Trp?Phe?Ala?Ser

180?????????????????185?????????????????190

Phe?Met?Ser?Ser?Tyr?Met?Ser?Met?Trp?Gln?Phe?Ala?Arg?Leu?Ala?Trp

195?????????????????200?????????????????205

Trp?Thr?Val?Val?Met?Gln?Leu?Leu?Gly?Ala?Pro?Met?Ala?Asn?Leu?Leu

210?????????????????215?????????????????220

Val?Phe?Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu?Phe?Tyr

225?????????????????230?????????????????235?????????????????240

Phe?Gly?Thr?Tyr?Met?Pro?His?Lys?Pro?Glu?Pro?Gly?Ala?Ala?Ser?Gly

245?????????????????250?????????????????255

Ser?Ser?Pro?Ala?Val?Met?Asn?Trp?Trp?Lys?Ser?Arg?Thr?Ser?Gln?Ala

260?????????????????265?????????????????270

Ser?Asp?Leu?Val?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?His?Trp

275?????????????????280?????????????????285

Glu?His?His?Arg?Trp?Pro?Phe?Ala?Pro?Trp?Trp?Glu?Leu?Pro?Asn?Cys

290?????????????????295?????????????????300

Arg?Arg?Leu?Ser?Gly?Arg?Gly?Leu?Val?Pro?Ala

305?????????????????310?????????????????315

<210>26

<211>1031

<212>DNA

＜213〉haematococcus pluvialis

<220>

<221>CDS

<222>(6)..(1031)

<223>

<400>26

gaagc?atg?cag?cta?gca?gcg?aca?gta?atg?ttg?gag?cag?ctt?acc?gga?agc??????50

Met?Gln?Leu?Ala?Ala?Thr?Val?Met?Leu?Glu?Gln?Leu?Thr?Gly?Ser

1???????????????5???????????????????10??????????????????15

gct?gag?gca?ctc?aag?gag?aag?gag?aag?gag?gtt?gca?ggc?agc?tct?gac????????98

Ala?Glu?Ala?Leu?Lys?Glu?Lys?Glu?Lys?Glu?Val?Ala?Gly?Ser?Ser?Asp

20??????????????????25??????????????????30

gtg?ttg?cgt?aca?tgg?gcg?acc?cag?tac?tcg?ctt?ccg?tca?gag?gag?tca???????146

Val?Leu?Arg?Thr?Trp?Ala?Thr?Gln?Tyr?Ser?Leu?Pro?Ser?Glu?Glu?Ser

35??????????????????40??????????????????45

gac?gcg?gcc?cgc?ccg?gga?ctg?aag?aat?gcc?tac?aag?cca?cca?cct?tcc???????194

Asp?Ala?Ala?Arg?Pro?Gly?Leu?Lys?Asn?Ala?Tyr?Lys?Pro?Pro?Pro?Ser

50??????????????????55??????????????????60

gac?aca?aag?ggc?atc?aca?atg?gcg?cta?gct?gtc?atc?ggc?tcc?tgg?gct???????242

Asp?Thr?Lys?Gly?Ile?Thr?Met?Ala?Leu?Ala?Val?Ile?Gly?Ser?Trp?Ala

65??????????????????70??????????????????75

gca?gtg?ttc?ctc?cac?gcc?att?ttt?caa?atc?aag?ctt?ccg?acc?tcc?ttg???????290

Ala?Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile?Lys?Leu?Pro?Thr?Ser?Leu

80??????????????????85??????????????????90??????????????????95

gac?cag?ctg?cac?tgg?ctg?ccc?gtg?tca?gat?gcc?aca?gct?cag?ctg?gtt???????338

Asp?Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Asp?Ala?Thr?Ala?Gln?Leu?Val

100?????????????????105?????????????????110

agc?ggc?agc?agc?agc?ctg?ctg?cac?atc?gtc?gta?gta?ttc?ttt?gtc?ctg???????386

Ser?Gly?Ser?Ser?Ser?Leu?Leu?His?Ile?Val?Val?Val?Phe?Phe?Val?Leu

115?????????????????120?????????????????125

gag?ttc?ctg?tac?aca?ggc?ctt?ttt?atc?acc?acg?cat?gat?gct?atg?cat???????434

Glu?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Thr?His?Asp?Ala?Met?His

130?????????????????135?????????????????140

ggc?acc?atc?gcc?atg?aga?aac?agg?cag?ctt?aat?gac?ttc?ttg?ggc?aga???????482

Gly?Thr?Ile?Ala?Met?Arg?Asn?Arg?Gln?Leu?Asn?Asp?Phe?Leu?Gly?Arg

145?????????????????150?????????????????155

gta?tgc?atc?tcc?ttg?tac?gcc?tgg?ttt?gat?tac?aac?atg?ctg?cac?cgc???????530

Val?Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp?Tyr?Asn?Met?Leu?His?Arg

160?????????????????165?????????????????170?????????????????175

aag?cat?tgg?gag?cac?cac?aac?cac?act?ggc?gag?gtg?ggc?aag?gac?cct???????578

Lys?His?Trp?Glu?His?His?Asn?His?Thr?Gly?Glu?Val?Gly?Lys?Asp?Pro

180?????????????????185?????????????????190

gac?ttc?cac?agg?gga?aac?cct?ggc?att?gtg?ccc?tgg?ttt?gcc?agc?ttc???????626

Asp?Phe?His?Arg?Gly?Asn?Pro?Gly?Ile?Val?Pro?Trp?Phe?Ala?Ser?Phe

195?????????????????200?????????????????205

atg?tcc?agc?tac?atg?tcg?atg?tgg?cag?ttt?gcg?cgc?ctc?gca?tgg?tgg???????674

Met?Ser?Ser?Tyr?Met?Ser?Met?Trp?Gln?Phe?Ala?Arg?Leu?Ala?Trp?Trp

210?????????????????215?????????????????220

acg?gtg?gtc?atg?cag?ctg?ctg?ggt?gcg?cca?atg?gcg?aac?ctg?ctg?gtg???????722

Thr?Val?Val?Met?Gln?Leu?Leu?Gly?Ala?Pro?Met?Ala?Asn?Leu?Leu?Val

225?????????????????230?????????????????235

ttc?atg?gcg?gcc?gcg?ccc?arc?ctg?tcc?gcc?ttc?cgc?ttg?ttc?tac?ttt???????770

Phe?Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu?Phe?Tyr?Phe

240?????????????????245?????????????????250?????????????????255

ggc?acg?tac?atg?ccc?cac?aag?cct?gag?cct?ggc?gcc?gcg?tca?ggc?tct???????818

Gly?Thr?Tyr?Met?Pro?His?Lys?Pro?Glu?Pro?Gly?Ala?Ala?Ser?Gly?Ser

260?????????????????265?????????????????270

tca?cca?gcc?gtc?atg?aac?tgg?tgg?aag?tcg?cgc?act?agc?cag?gcg?tcc???????866

Ser?Pro?Ala?Val?Met?Asn?Trp?Trp?Lys?Ser?Arg?Thr?Ser?Gln?Ala?Ser

275?????????????????280?????????????????285

gac?ctg?gtc?agc?ttt?ctg?acc?tgc?tac?cac?ttc?gac?ctg?cac?tgg?gag???????914

Asp?Leu?Val?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?His?Trp?Glu

290?????????????????295?????????????????300

cac?cac?cgc?tgg?ccc?ttt?gcc?ccc?tgg?tgg?gag?ctg?ccc?aac?tgc?cgc???????962

His?His?Arg?Trp?Pro?Phe?Ala?Pro?Trp?Trp?Glu?Leu?Pro?Asn?Cys?Arg

305?????????????????310?????????????????315

cgc?ctg?tct?ggc?cga?ggt?ctg?gtt?cct?gcc?gag?caa?aaa?ctc?atc?tca??????1010

Arg?Leu?Ser?Gly?Arg?Gly?Leu?Val?Pro?Ala?Glu?Gln?Lys?Leu?Ile?Ser

320?????????????????325?????????????????330?????????????????335

gaa?gag?gat?ctg?aat?agc?tag??????????????????????????????????????????1031

Glu?Glu?Asp?Leu?Asn?Ser

340

<210>27

<211>341

<212>PRT

＜213〉haematococcus pluvialis

<400>27

Met?Gln?Leu?Ala?Ala?Thr?Val?Met?Leu?Glu?Gln?Leu?Thr?Gly?Ser?Ala

1???????????????5???????????????????10??????????????????15

Glu?Ala?Leu?Lys?Glu?Lys?Glu?Lys?Glu?Val?Ala?Gly?Ser?Ser?Asp?Val

20??????????????????25??????????????????30

Leu?Arg?Thr?Trp?Ala?Thr?Gln?Tyr?Ser?Leu?Pro?Ser?Glu?Glu?Ser?Asp

35??????????????????40??????????????????45

Ala?Ala?Arg?Pro?Gly?Leu?Lys?Asn?Ala?Tyr?Lys?Pro?Pro?Pro?Ser?Asp

50??????????????????55??????????????????60

Thr?Lys?Gly?Ile?Thr?Met?Ala?Leu?Ala?Val?Ile?Gly?Ser?Trp?Ala?Ala

65??????????????????70??????????????????75??????????????????80

Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile?Lys?Leu?Pro?Thr?Ser?Leu?Asp

85??????????????????90??????????????????95

Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Asp?Ala?Thr?Ala?Gln?Leu?Val?Ser

100?????????????????105?????????????????110

Gly?Ser?Ser?Ser?Leu?Leu?His?Ile?Val?Val?Val?Phe?Phe?Val?Leu?Glu

115?????????????????120?????????????????125

Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Thr?His?Asp?Ala?Met?His?Gly

130?????????????????135?????????????????140

Thr?Ile?Ala?Met?Arg?Asn?Arg?Gln?Leu?Asn?Asp?Phe?Leu?Gly?Arg?Val

145?????????????????150?????????????????155?????????????????160

Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp?Tyr?Asn?Met?Leu?His?Arg?Lys

165?????????????????170?????????????????175

His?Trp?Glu?His?His?Asn?His?Thr?Gly?Glu?Val?Gly?Lys?Asp?Pro?Asp

180?????????????????185?????????????????190

Phe?His?Arg?Gly?Asn?Pro?Gly?Ile?Val?Pro?Trp?Phe?Ala?Ser?Phe?Met

195?????????????????200?????????????????205

Ser?Ser?Tyr?Met?Ser?Met?Trp?Gln?Phe?Ala?Arg?Leu?Ala?Trp?Trp?Thr

210?????????????????215?????????????????220

Val?Val?Met?Gln?Leu?Leu?Gly?Ala?Pro?Met?Ala?Asn?Leu?Leu?Val?Phe

225?????????????????230?????????????????235?????????????????240

Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu?Phe?Tyr?Phe?Gly

245?????????????????250?????????????????255

Thr?Tyr?Met?Pro?His?Lys?Pro?Glu?Pro?Gly?Ala?Ala?Ser?Gly?Ser?Ser

260?????????????????265?????????????????270

Pro?Ala?Val?Met?Asn?Trp?Trp?Lys?Ser?Arg?Thr?Ser?Gln?Ala?Ser?Asp

275?????????????????280?????????????????285

Leu?Val?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?His?Trp?Glu?His

290?????????????????295?????????????????300

His?Arg?Trp?Pro?Phe?Ala?Pro?Trp?Trp?Glu?Leu?Pro?Asn?Cys?Arg?Arg

305?????????????????310?????????????????315?????????????????320

Leu?Ser?Gly?Arg?Gly?Leu?Val?Pro?Ala?Glu?Gln?Lys?Leu?Ile?Ser?Glu

325?????????????????330?????????????????335

Glu?Asp?Leu?Asn?Ser

340

<210>28

<211>777

<212>DNA

＜213〉mouseearcress (Arabidopsis thaliana)

<220>

＜221〉promoter

<222>(1)..(777)

<223>

<400>28

gagctcactc?actgatttcc?attgcttgaa?aattgatgat?gaactaagat?caatccatgt?????60

tagtttcaaa?acaacagtaa?ctgtggccaa?cttagttttg?aaacaacact?aactggtcga????120

agcaaaaaga?aaaaagagtt?tcatcatata?tctgatttga?tggactgttt?ggagttagga????180

ccaaacatta?tctacaaaca?aagacttttc?tcctaacttg?tgattccttc?ttaaacccta????240

ggggtaatat?tctattttcc?aaggatcttt?agttaaaggc?aaatccggga?aattattgta????300

atcatttggg?gaaacatata?aaagatttga?gttagatgga?agtgacgatt?aatccaaaca????360

tatatatctc?tttcttctta?tttcccaaat?taacagacaa?aagtagaata?ttggctttta????420

acaccaatat?aaaaacttgc?ttcacaccta?aacacttttg?tttactttag?ggtaagtgca????480

aaaagccaac?caaatccacc?tgcactgatt?tgacgtttac?aaacgccgtt?aagtcgatgt????540

ccgttgattt?aaacagtgtc?ttgtaattaa?aaaaatcagt?ttacataaat?ggaaaattta????600

tcacttagtt?ttcatcaact?tctgaactta?cctttcatgg?attaggcaat?actttccatt????660

tttagtaact?caagtggacc?ctttacttct?tcaactccat?ctctctcttt?ctatttcact????720

tctttcttct?cattatatct?cttgtcctct?ccaccaaatc?tcttcaacaa?aaagctt???????777

<210>29

<211>22

<212>DNA

＜213〉synthesize

<220>

＜221〉primer _ combination

<222>(1)..(22)

<223>

<400>29

gcaagctcga?cagctacaaa?cc??????????????????????????????????22

<210>30

<211>24

<212>DNA

＜213〉synthesize

<220>

＜221〉primer _ combination

<222>(1)..(24)

<223>

<400>30

gaagcatgca?gctagcagcg?acag????????????????????????????????24

<210>31

<211>30

<212>DNA

＜213〉synthesize

<220>

＜221〉primer _ combination

<222>(1)..(30)

<223>

<400>31

tgcatgctag?aggcactcaa?ggagaaggag????????????????????????????????????30

<210>32

<211>59

<212>DNA

＜213〉synthesize

<220>

＜221〉primer _ combination

<222>(1)..(59)

<223>

<400>32

ctagctattc?agatcctctt?ctgagatgag?tttttgctcg?gcaggaacca?gacctcggc???59

<210>33

<211>28

<212>DNA

＜213〉synthesize

<220>

＜221〉primer _ combination

<222>(1)..(28)

<223>

<400>33

gagctcactc?actgatttcc?attgcttg???????????????????????????????????28

<210>34

<211>37

<212>DNA

＜213〉synthesize

<220>

＜221〉primer _ combination

<222>(1)..(37)

<223>

<400>34

cgccgttaag?tcgatgtccg?ttgatttaaa?cagtgtc?????????????????????????37

<210>35

<211>34

<212>DNA

＜213〉synthesize

<220>

＜221〉primer _ combination

<222>(1)..(34)

<223>

<400>35

atcaacggac?atcgacttaa?cggcgtttgt?aaac???????????????????????????34

<210>36

<211>25

<212>DNA

＜213〉synthesize

<220>

＜221〉primer _ combination

<222>(1)..(25)

<223>

<400>36

taagcttttt?gttgaagaga?tttgg?????????????????????????????????????25

<210>37

<211>212

<212>DNA

＜213〉composition sequence

<220>

＜221〉introne

<222>(1)..(212)

<223>

<400>37

gtcgactacg?taagtttctg?cttctacctt?tgatatatat?ataataatta?tcattaatta????60

gtagtaatat?aatatttcaa?atattttttt?caaaataaaa?gaatgtagta?tatagcaatt????120

gcttttctgt?agtttataag?tgtgtatatt?ttaatttata?acttttctaa?tatatgacca????180

aaatttgttg?atgtgcaggt?atcaccggat?cc??????????????????????????????????212

<210>38

<211>1830

<212>DNA

＜213〉marigold (Tagetes erecta)

<220>

<221>CDS

<222>(141)..(1691)

<223>

<400>38

ggcacgaggc?aaagcaaagg?ttgtttgttg?ttgttgttga?gagacactcc?aatccaaaca?????60

gatacaaggc?gtgactggat?atttctctct?cgttcctaac?aacagcaacg?aagaagaaaa????120

agaatcatta?ctaacaatca?atg?agt?atg?aga?gct?gga?cac?atg?acg?gca?aca????173

Met?Ser?Met?Arg?Ala?Gly?His?Met?Thr?Ala?Thr

1???????????????5???????????????????10

atg?gcg?gct?ttt?aca?tgc?cct?agg?ttt?atg?act?agc?atc?aga?tac?acg??????221

Met?Ala?Ala?Phe?Thr?Cys?Pro?Arg?Phe?Met?Thr?Ser?Ile?Arg?Tyr?Thr

15??????????????????20??????????????????25

aag?caa?att?aag?tgc?aac?gct?gct?aaa?agc?cag?cta?gtc?gtt?aaa?caa??????269

Lys?Gln?Ile?Lys?Cys?Asn?Ala?Ala?Lys?Ser?Gln?Leu?Val?Val?Lys?Gln

30??????????????????35??????????????????40

gag?att?gag?gag?gaa?gaa?gat?tat?gtg?aaa?gcc?ggt?gga?tcg?gag?ctg??????317

Glu?Ile?Glu?Glu?Glu?Glu?Asp?Tyr?Val?Lys?Ala?Gly?Gly?Ser?Glu?Leu

45??????????????????50??????????????????55

ctt?ttt?gtt?caa?atg?caa?cag?aat?aag?tcc?atg?gat?gca?cag?tct?agc??????365

Leu?Phe?Val?Gln?Met?Gln?Gln?Asn?Lys?Ser?Met?Asp?Ala?Gln?Ser?Ser

60??????????????????65??????????????????70??????????????????75

cta?tcc?caa?aag?ctc?cca?agg?gta?cca?ata?gga?gga?gga?gga?gac?agt??????413

Leu?Ser?Gln?Lys?Leu?Pro?Arg?Val?Pro?Ile?Gly?Gly?Gly?Gly?Asp?Ser

80??????????????????85??????????????????90

aac?tgt?ata?ctg?gat?ttg?gtt?gta?att?ggt?tgt?ggt?cct?gct?ggc?ctt??????461

Asn?Cys?Ile?Leu?Asp?Leu?Val?Val?Ile?Gly?Cys?Gly?Pro?Ala?Gly?Leu

95??????????????????100?????????????????105

gct?ctt?gct?gga?gaa?tca?gcc?aag?cta?ggc?ttg?aat?gtc?gca?ctt?atc??????509

Ala?Leu?Ala?Gly?Glu?Ser?Ala?Lys?Leu?Gly?Leu?Asn?Val?Ala?Leu?Ile

110?????????????????115?????????????????120

ggc?cct?gat?ctt?cct?ttt?aca?aat?aac?tat?ggt?gtt?tgg?gag?gat?gaa??????557

Gly?Pro?Asp?Leu?Pro?Phe?Thr?Asn?Asn?Tyr?Gly?Val?Trp?Glu?Asp?Glu

125?????????????????130?????????????????135

ttt?ata?ggt?ctt?gga?ctt?gag?ggc?tgt?att?gaa?cat?gtt?tgg?cga?gat??????605

Phe?Ile?Gly?Leu?Gly?Leu?Glu?Gly?Cys?Ile?Glu?His?Val?Trp?Arg?Asp

140?????????????????145?????????????????150?????????????????155

act?gta?gta?tat?ctt?gat?gac?aac?gat?ccc?att?ctc?ata?ggt?cgt?gcc????653

Thr?Val?Val?Tyr?Leu?Asp?Asp?Asn?Asp?Pro?Ile?Leu?Ile?Gly?Arg?Ala

160?????????????????165?????????????????170

tat?gga?cga?gtt?agt?cgt?gat?tta?ctt?cac?gag?gag?ttg?ttg?act?agg????701

Tyr?Gly?Arg?Val?Ser?Arg?Asp?Leu?Leu?His?Glu?Glu?Leu?Leu?Thr?Arg

175?????????????????180?????????????????185

tgc?atg?gag?tca?ggc?gtt?tca?tat?ctg?agc?tcc?aaa?gtg?gaa?cgg?att????749

Cys?Met?Glu?Ser?Gly?Val?Ser?Tyr?Leu?Ser?Ser?Lys?Val?Glu?Arg?Ile

190?????????????????195?????????????????200

act?gaa?gct?cca?aat?ggc?cta?agt?ctc?ata?gag?tgt?gaa?ggc?aat?atc????797

Thr?Glu?Ala?Pro?Asn?Gly?Leu?Ser?Leu?Ile?Glu?Cys?Glu?Gly?Asn?Ile

205?????????????????210?????????????????215

aca?att?cca?tgc?agg?ctt?gct?act?gtc?gct?tct?gga?gca?gct?tct?gga????845

Thr?Ile?Pro?Cys?Arg?Leu?Ala?Thr?Val?Ala?Ser?Gly?Ala?Ala?Ser?Gly

220?????????????????225?????????????????230?????????????????235

aaa?ctt?ttg?cag?tat?gaa?ctt?ggc?ggt?ccc?cgt?gtt?tgc?gtt?caa?aca????893

Lys?Leu?Leu?Gln?Tyr?Glu?Leu?Gly?Gly?Pro?Arg?Val?Cys?Val?Gln?Thr

240?????????????????245?????????????????250

gct?tat?ggt?ata?gag?gtt?gag?gtt?gaa?agc?ata?ccc?tat?gat?cca?agc????941

Ala?Tyr?Gly?Ile?Glu?Val?Glu?Val?Glu?Ser?Ile?Pro?Tyr?Asp?Pro?Ser

255?????????????????260?????????????????265

cta?atg?gtt?ttc?atg?gat?tat?aga?gac?tac?acc?aaa?cat?aaa?tct?caa????989

Leu?Met?Val?Phe?Met?Asp?Tyr?Arg?Asp?Tyr?Thr?Lys?His?Lys?Ser?Gln

270?????????????????275?????????????????280

tca?cta?gaa?gca?caa?tat?cca?aca?ttt?ttg?tat?gtc?atg?cca?atg?tct????1037

Ser?Leu?Glu?Ala?Gln?Tyr?Pro?Thr?Phe?Leu?Tyr?Val?Met?Pro?Met?Ser

285?????????????????290?????????????????295

cca?act?aaa?gta?ttc?ttt?gag?gaa?act?tgt?ttg?gct?tca?aaa?gag?gcc????1085

Pro?Thr?Lys?Val?Phe?Phe?Glu?Glu?Thr?Cys?Leu?Ala?Ser?Lys?Glu?Ala

300?????????????????305?????????????????310?????????????????315

atg?cct?ttt?gag?tta?ttg?aag?aca?aaa?ctc?atg?tca?aga?tta?aag?act????1133

Met?Pro?Phe?Glu?Leu?Leu?Lys?Thr?Lys?Leu?Met?Ser?Arg?Leu?Lys?Thr

320?????????????????325?????????????????330

atg?ggg?atc?cga?ata?acc?aaa?act?tat?gaa?gag?gaa?tgg?tca?tat?att????1181

Met?Gly?Ile?Arg?Ile?Thr?Lys?Thr?Tyr?Glu?Glu?Glu?Trp?Ser?Tyr?Ile

335?????????????????340?????????????????345

cca?gta?ggt?gga?tcc?tta?cca?aat?acc?gag?caa?aag?aac?ctt?gca?ttt????1229

Pro?Val?Gly?Gly?Ser?Leu?Pro?Asn?Thr?Glu?Gln?Lys?Asn?Leu?Ala?Phe

350?????????????????355?????????????????360

ggt?gct?gct?gct?agc?atg?gtg?cat?cca?gcc?aca?gga?tat?tcg?gtt?gta????1277

Gly?Ala?Ala?Ala?Ser?Met?Val?His?Pro?Ala?Thr?Gly?Tyr?Ser?Val?Val

365?????????????????370?????????????????375

aga?tca?ctg?tca?gaa?gct?cct?aat?tat?gca?gca?gta?att?gca?aag?att????1325

Arg?Ser?Leu?Ser?Glu?Ala?Pro?Asn?Tyr?Ala?Ala?Val?Ile?Ala?Lys?Ile

380?????????????????385?????????????????390?????????????????395

tta?ggg?aaa?gga?aat?tca?aaa?cag?atg?ctt?gat?cat?gga?aga?tac?aca????1373

Leu?Gly?Lys?Gly?Asn?Ser?Lys?Gln?Met?Leu?Asp?His?Gly?Arg?Tyr?Thr

400?????????????????405?????????????????410

acc?aac?atc?tca?aag?caa?gct?tgg?gaa?aca?ctt?tgg?ccc?ctt?gaa?agg????1421

Thr?Asn?Ile?Ser?Lys?Gln?Ala?Trp?Glu?Thr?Leu?Trp?Pro?Leu?Glu?Arg

415?????????????????420?????????????????425

aaa?aga?cag?aga?gca?ttc?ttt?ctc?ttt?gga?tta?gca?ctg?att?gtc?cag????1469

Lys?Arg?Gln?Arg?Ala?Phe?Phe?Leu?Phe?Gly?Leu?Ala?Leu?Ile?Val?Gln

430?????????????????435?????????????????440

atg?gat?att?gag?ggg?acc?cgc?aca?ttc?ttc?cgg?act?ttc?ttc?cgc?ttg????1517

Met?Asp?Ile?Glu?Gly?Thr?Arg?Thr?Phe?Phe?Arg?Thr?Phe?Phe?Arg?Leu

445?????????????????450?????????????????455

ccc?aca?tgg?atg?tgg?tgg?ggg?ttt?ctt?gga?tct?tcg?tta?tca?tca?act????1565

Pro?Thr?Trp?Met?Trp?Trp?Gly?Phe?Leu?Gly?Ser?Ser?Leu?Ser?Ser?Thr

460?????????????????465?????????????????470?????????????????475

gac?ttg?ata?ata?ttt?gcg?ttt?tac?atg?ttt?atc?ata?gca?ccg?cat?agc????1613

Asp?Leu?Ile?Ile?Phe?Ala?Phe?Tyr?Met?Phe?Ile?Ile?Ala?Pro?His?Ser

480?????????????????485?????????????????490

ctg?aga?atg?ggt?ctg?gtt?aga?cat?ttg?ctt?tct?gac?ccg?aca?gga?gga????1661

Leu?Arg?Met?Gly?Leu?Val?Arg?His?Leu?Leu?Ser?Asp?Pro?Thr?Gly?Gly

495?????????????????500?????????????????505

aca?atg?tta?aaa?gcg?tat?ctc?acg?ata?taa?ataactctag?tcgcgatcag??????1711

Thr?Met?Leu?Lys?Ala?Tyr?Leu?Thr?Ile

510?????????????????515

tttagattat?aggcacatct?tgcatatata?tatgtataaa?ccttatgtgt?gctgtatcct??1771

tacatcaaca?cagtcattaa?ttgtatttct?tggggtaatg?ctgatgaagt?attttctgg???1830

<210>39

<211>516

<212>PRT

＜213〉marigold

<400>39

Met?Ser?Met?Arg?Ala?Gly?His?Met?Thr?Ala?Thr?Met?Ala?Ala?Phe?Thr

1???????????????5???????????????????10??????????????????15

Cys?Pro?Arg?Phe?Met?Thr?Ser?Ile?Arg?Tyr?Thr?Lys?Gln?Ile?Lys?Cys

20??????????????????25??????????????????30

Asn?Ala?Ala?Lys?Ser?Gln?Leu?Val?Val?Lys?Gln?Glu?Ile?Glu?Glu?Glu

35??????????????????40??????????????????45

Glu?Asp?Tyr?Val?Lys?Ala?Gly?Gly?Ser?Glu?Leu?Leu?Phe?Val?Gln?Met

50??????????????????55??????????????????60

Gln?Gln?Asn?Lys?Ser?Met?Asp?Ala?Gln?Ser?Ser?Leu?Ser?Gln?Lys?Leu

65??????????????????70??????????????????75??????????????????80

Pro?Arg?Val?Pro?Ile?Gly?Gly?Gly?Gly?Asp?Ser?Asn?Cys?Ile?Leu?Asp

85??????????????????90??????????????????95

Leu?Val?Val?Ile?Gly?Cys?Gly?Pro?Ala?Gly?Leu?Ala?Leu?Ala?Gly?Glu

100?????????????????105?????????????????110

Ser?Ala?Lys?Leu?Gly?Leu?Asn?Val?Ala?Leu?Ile?Gly?Pro?Asp?Leu?Pro

115?????????????????120?????????????????125

Phe?Thr?Asn?Asn?Tyr?Gly?Val?Trp?Glu?Asp?Glu?Phe?Ile?Gly?Leu?Gly

130?????????????????135?????????????????140

Leu?Glu?Gly?Cys?Ile?Glu?His?Val?Trp?Arg?Asp?Thr?Val?Val?Tyr?Leu

145?????????????????150?????????????????155?????????????????160

Asp?Asp?Asn?Asp?Pro?Ile?Leu?Ile?Gly?Arg?Ala?Tyr?Gly?Arg?Val?Ser

165?????????????????170?????????????????175

Arg?Asp?Leu?Leu?His?Glu?Glu?Leu?Leu?Thr?Arg?Cys?Met?Glu?Ser?Gly

180?????????????????185?????????????????190

Val?Ser?Tyr?Leu?Ser?Ser?Lys?Val?Glu?Arg?Ile?Thr?Glu?Ala?Pro?Asn

195?????????????????200??????????????????205

Gly?Leu?Ser?Leu?Ile?Glu?Cys?Glu?Gly?Asn?Ile?Thr?Ile?Pro?Cys?Arg

210?????????????????215?????????????????220

Leu?Ala?Thr?Val?Ala?Ser?Gly?Ala?Ala?Ser?Gly?Lys?Leu?Leu?Gln?Tyr

225?????????????????230?????????????????235?????????????????240

Glu?Leu?Gly?Gly?Pro?Arg?Val?Cys?Val?Gln?Thr?Ala?Tyr?Gly?Ile?Glu

245?????????????????250?????????????????255

Val?Glu?Val?Glu?Ser?Ile?Pro?Tyr?Asp?Pro?Ser?Leu?Met?Val?Phe?Met

260?????????????????265?????????????????270

Asp?Tyr?Arg?Asp?Tyr?Thr?Lys?His?Lys?Ser?Gln?Ser?Leu?Glu?Ala?Gln

275?????????????????280?????????????????285

Tyr?Pro?Thr?Phe?Leu?Tyr?Val?Met?Pro?Met?Ser?Pro?Thr?Lys?Val?Phe

290?????????????????295?????????????????300

Phe?Glu?Glu?Thr?Cys?Leu?Ala?Ser?Lys?Glu?Ala?Met?Pro?Phe?Glu?Leu

305?????????????????310?????????????????315?????????????????320

Leu?Lys?Thr?Lys?Leu?Met?Ser?Arg?Leu?Lys?Thr?Met?Gly?Ile?Arg?Ile

325?????????????????330?????????????????335

Thr?Lys?Thr?Tyr?Glu?Glu?Glu?Trp?Ser?Tyr?Ile?Pro?Val?Gly?Gly?Ser

340?????????????????345?????????????????350

Leu?Pro?Asn?Thr?Glu?Gln?Lys?Asn?Leu?Ala?Phe?Gly?Ala?Ala?Ala?Ser

355?????????????????360?????????????????365

Met?Val?His?Pro?Ala?Thr?Gly?Tyr?Ser?Val?Val?Arg?Ser?Leu?Ser?Glu

370?????????????????375?????????????????380

Ala?Pro?Asn?Tyr?Ala?Ala?Val?Ile?Ala?Lys?Ile?Leu?Gly?Lys?Gly?Asn

385?????????????????390??????????????????395????????????????400

Ser?Lys?Gln?Met?Leu?Asp?His?Gly?Arg?Tyr?Thr?Thr?Asn?Ile?Ser?Lys

405?????????????????410?????????????????415

Gln?Ala?Trp?Glu?Thr?Leu?Trp?Pro?Leu?Glu?Arg?Lys?Arg?Gln?Arg?Ala

420?????????????????425?????????????????430

Phe?Phe?Leu?Phe?Gly?Leu?Ala?Leu?Ile?Val?Gln?Met?Asp?Ile?Glu?Gly

435?????????????????440?????????????????445

Thr?Arg?Thr?Phe?Phe?Arg?Thr?Phe?Phe?Arg?Leu?Pro?Thr?Trp?Met?Trp

450?????????????????455?????????????????460

Trp?Gly?Phe?Leu?Gly?Ser?Ser?Leu?Ser?Ser?Thr?Asp?Leu?Ile?Ile?Phe

465?????????????????470?????????????????475?????????????????480

Ala?Phe?Tyr?Met?Phe?Ile?Ile?Ala?Pro?His?Ser?Leu?Arg?Met?Gly?Leu

485?????????????????490?????????????????495

Val?Arg?His?Leu?Leu?Ser?Asp?Pro?Thr?Gly?Gly?Thr?Met?Leu?Lys?Ala

500?????????????????505?????????????????510

Tyr?Leu?Thr?Ile

515

<210>40

<211>445

<212>DNA

＜213〉marigold

<220>

＜221〉have a mind to fragment

<222>(1)..(445)

<223>

<400>40

aagcttgcac?gaggcaaagc?aaaggttgtt?tgttgttgtt?gttgagagac?actccaatcc?????60

aaacagatac?aaggcgtgac?tggatatttc?tctctcgttc?ctaacaacag?caacgaagaa????120

gaaaaagaat?cattactaac?aatcaatgag?tatgagagct?ggacacatga?cggcaacaat????180

ggcggctttt?acatgcccta?ggtttatgac?tagcatcaga?tacacgaagc?aaattaagtg????240

caacgctgct?aaaagccagc?tagtcgttaa?acaagagatt?gaggaggaag?aagattatgt????300

gaaagccggt?ggatcggagc?tgctttttgt?tcaaatgcaa?cagaataagt?ccatggatgc????360

acagtctagc?ctatcccaaa?agctcccaag?ggtaccaata?ggaggaggag?gagacagtaa????420

ctgtatactg?gatttggttg?tcgac??????????????????????????????????????????445

<210>41

<211>446

<212>DNA

＜213〉marigold

<220>

＜221〉antisense fragment

<222>(1)..(446)

<223>

<400>41

gaattcgcac?gaggcaaagc?aaaggttgtt?tgttgttgtt?gttgagagac?actccaatcc?????60

aaacagatac?aaggcgtgac?tggatatttc?tctctcgttc?ctaacaacag?caacgaagaa????120

gaaaaagaat?cattactaac?aatcaatgag?tatgagagct?ggacacatga?cggcaacaat????180

ggcggctttt?acatgcccta?ggtttatgac?tagcatcaga?tacacgaagc?aaattaagtg????240

caacgctgct?aaaagccagc?tagtcgttaa?acaagagatt?gaggaggaag?aagattatgt????300

gaaagccggt?ggatcggagc?tgctttttgt?tcaaatgcaa?cagaataagt?ccatggatgc????360

acagtctagc?ctatcccaaa?agctcccaag?ggtaccaata?ggaggaggag?gagacagtaa????420

ctgtatactg?gatttggttg?gatcct?????????????????????????????????????????446

<210>42

<211>393

<212>DNA

＜213〉marigold

<220>

＜221〉have a mind to fragment

<222>(1)..(393)

<223>

<400>42

aagctttgga?ttagcactga?ttgtccagat?ggatattgag?gggacccgca?cattcttccg?????60

gactttcttc?cgcttgccca?catggatgtg?gtgggggttt?cttggatctt?cgttatcatc????120

aactgacttg?ataatatttg?cgttttacat?gtttatcata?gcaccgcata?gcctgagaat????180

gggtctggtt?agacatttgc?tttctgaccc?gacaggagga?acaatgttaa?aagcgtatct????240

cacgatataa?ataactctag?tcgcgatcag?tttagattat?aggcacatct?tgcatatata????300

tatgtataaa?ccttatgtgt?gctgtatcct?tacatcaaca?cagtcattaa?ttgtatttct????360

tggggtaatg?ctgatgaagt?attttctgtc?gac?????????????????????????????????393

<210>43

<211>397

<212>DNA

＜213〉marigold

<220>

＜221〉antisense fragment

<222>(1)..(397)

<223>

<400>43

gaattctctt?tggattagca?ctgattgtcc?agatggatat?tgaggggacc?cgcacattct?????60

tccggacttt?cttccgcttg?cccacatgga?tgtggtgggg?gtttcttgga?tcttcgttat????120

catcaactga?cttgataata?tttgcgtttt?acatgtttat?catagcaccg?catagcctga????180

gaatgggtct?ggttagacat?ttgctttctg?acccgacagg?aggaacaatg?ttaaaagcgt????240

atctcacgat?ataaataact?ctagtcgcga?tcagtttaga?ttataggcac?atcttgcata????300

tatatatgta?taaaccttat?gtgtgctgta?tccttacatc?aacacagtca?ttaattgtat????360

ttcttggggt?aatgctgatg?aagtattttc?tggatcc?????????????????????????????397

<210>44

<211>1537

<212>DNA

<213>-

<220>

＜221〉promoter

<222>(1)..(1537)

<223>

<400>44

gagctctaca?aattagggtt?actttattca?ttttcatcca?ttctctttat?tgttaaattt?????60

tgtacattta?ttcaataata?ttatatgttt?attacaaatt?ctcactttct?tattcatacc????120

tattcactca?agcctttacc?atcttccttt?tctatttcaa?tactatttct?acttcatttt????180

tcacgttttt?aacatctttc?tttatttctt?gtccacttcg?tttagggatg?cctaatgtcc????240

caaatttcat?ctctcgtagt?aacacaaaac?caatgtaatg?ctacttctct?ctacattttt????300

aatacaaata?aagtgaaaca?aaatatctat?aaataaacaa?atatatatat?tttgttagac????360

gctgtctcaa?cccatcaatt?aaaaaatttt?gttatatttc?tactttacct?actaaatttg????420

tttctcatat?ttacctttta?acccccacaa?aaaaaaatta?taaaaaagaa?agaaaaaagc????480

taaaccctat?ttaaatagct?aactataaga?tcttaaaatt?atcctcatca?gtgtatagtt????540

taattggtta?ttaacttata?acattatata?tctatgacat?atactctctc?ctagctattt????600

ctcacatttt?ttaacttaag?aaaatagtca?taacatagtc?taaaattcaa?acatccacat????660

gctctaattt?gattaacaaa?aagttagaaa?tatttattta?aataaaaaag?actaataaat????720

atataaaatg?aatgttcata?cgcagaccca?tttagagatg?agtatgcttt?cacatgctga????780

gattattttc?aaaactaagg?ttgtagcaat?attaaatcaa?taaaattatt?ataaataaca????840

aaattaacct?gctcgtgttt?gctgtatatg?ggaggctaca?aaataaatta?aactaaagat????900

gattatgttt?tagacatttt?ttctatctgt?attagtttat?acatattaat?tcaggagctg????960

cacaacccaa?ttctattttc?gttccttggt?ggctgggttt?ctcacaaggt?tcaatagtca????1020

atattaggtt?ttattggact?tttaatagta?tcaaacaaat?ctatgtgtga?acttaaaaat????1080

tgtattaaat?atttagggta?acctgttgcc?gtttttagaa?taatgtttct?tcttaataca????1140

cgaaagcgta?ttgtgtattc?attcatttgg?cgcctcacat?gcttcggttg?gctcgcttta????1200

gtctctgcct?tctttgtata?ttgtactccc?cctcttccta?tgccacgtgt?tctgagctta????1260

acaagccacg?ttgcgtgcca?ttgccaaaca?agtcatttta?acttcacaag?gtccgatttg????1320

acctccaaaa?caacgacaag?tttccgaaca?gtcgcgaaga?tcaagggtat?aatcgtcttt????1380

ttgaattcta?tttctcttta?tttaatagtc?cctctcgtgt?gatagttttt?aaaagatttt????1440

taaaacgtag?ctgctgttta?agtaaatccc?agtccttcag?tttgtgcttt?tgtgtgtttt????1500

gtttctctga?tttacggaat?ttggaaataa?taagctt?????????????????????????????1537

<210>45

<211>734

<212>DNA

＜213〉composition sequence

<220>

＜221〉variation

<222>(1)..(734)

<223>

<400>45

ctaacaatca?atgagtagag?agctggacac?atgacggcaa?caatggcggc?ttttacatgc?????60

cctaggttta?tgactagcat?cagatacacg?aagcaaatta?agtgcaacgc?tgctaaaagc????120

cagctagtcg?ttaaacaaga?gattgaggag?gaagaagatt?atgtgaaagc?cggtggatcg????180

gagctgcttt?ttgttcaaat?gcaacagaat?aagtccatgg?atgcacagtc?tagcctatcc????240

caaaaggtca?ctccagactt?aattgcttat?aaataaataa?atatgttttt?taggaataat????300

gatatttaga?tagattagct?atcacctgtg?ctgtggtgtg?cagctcccaa?gggtcttacc????360

gatagtaaaa?tcgttagtta?tgattaatac?ttgggaggtg?ggggattata?ggctttgttg????420

tgagaatgtt?gagaaagagg?tttgacaaat?cggtgtttga?atgaggttaa?atggagttta????480

attaaaataa?agagaagaga?aagattaaga?gggtgatggg?gatattaaag?acggscaata????540

tagtgatgcc?acgtagaaaa?aggtaagtga?aaacatacaa?cgtggcttta?aaagatggct????600

tggctgctaa?tcaactcaac?tcaactcata?tcctatccat?tcaaattcaa?ttcaattcta????660

ttgaatgcaa?agcaaagcaa?aggttgtttg?ttgttgttgt?tgagagacac?tccaatccaa????720

acagatacaa?ggcg??????????????????????????????????????????????????????734

<210>46

<211>280

<212>DNA

＜213〉composition sequence

<220>

＜221〉variation

<222>(1)..(280)

<223>

<400>46

gtcgagtatg?gagttcaatt?aaaataaaga?gaagaraaag?attaagaggg?tgatggggat?????60

attaaagacg?gccaatrtag?tgatgccacg?taagaaaaag?gtaagtgaaa?acatacaacg????120

tggctttaaa?agatggcttg?gctgctaatc?aactcaactc?aactcatatc?ctatccattc????180

aaattcaatt?caattctatt?gaatgcaaag?caaagcaaag?caaaggttgt?ttgttgttgt????240

tgttgagaga?cactccaatc?caaacagata?caaggcgtga??????????????????????????280

<210>47

<211>358

<212>DNA

＜213〉marigold

<220>

＜221〉have a mind to promoter

<222>(1)..(358)

<223>

<400>47

aagcttaccg?atagtaaaat?cgttagttat?gattaatact?tgggaggtgg?gggattatag?????60

gctttgttgt?gagaatgttg?agaaagaggt?ttgacaaatc?ggtgtttgaa?tgaggttaaa????120

tggagtttaa?ttaaaataaa?gagaagagaa?agattaagag?ggtgatgggg?atattaaaga????180

cggccaatat?agtgatgcca?cgtagaaaaa?ggtaagtgaa?aacatacaac?gtggctttaa????240

aagatggctt?ggctgctaat?caactcaact?caactcatat?cctatccatt?caaattcaat????300

tcaattctat?tgaatgcaaa?gcaaagcaaa?gcaaaggttg?tttgttgttg?ttgtcgac??????358

<210>48

<211>361

<212>DNA

＜213〉marigold

<220>

＜221〉antisense promoter

<222>(1)..(361)

<223>

<400>48

ctcgagctta?ccgatagtaa?aatcgttagt?tatgattaat?acttgggagg?tgggggatta?????60

taggctttgt?tgtgagaatg?ttgagaaaga?ggtttgacaa?atcggtgttt?gaatgaggtt????120

aaatggagtt?taattaaaat?aaagagaaga?gaaagattaa?gagggtgatg?gggatattaa????180

agacggccaa?tatagtgatg?ccacgtagaa?aaaggtaagt?gaaaacatac?aacgtggctt????240

taaaagatgg?cttggctgct?aatcaactca?actcaactca?tatcctatcc?attcaaattc????300

aattcaattc?tattgaatgc?aaagcaaagc?aaagcaaagg?ttgtttgttg?ttgttggatc????360

c????????????????????????????????????????????????????????????????????361

<210>49

<211>28

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(28)

<223>

<400>49

gagctcactc?actgatttcc?attgcttg??????????????????????????????????28

<210>50

<211>37

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(37)

<223>

<400>50

cgccgttaag?tcgatgtccg?ttgatttaaa?cagtgtc???????????????????????37

<210>51

<211>34

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(34)

<223>

<400>51

atcaacggac?atcgacttaa?cggcgtttgt?aaac??????????????????????????34

<210>52

<211>25

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(25)

<223>

<400>52

taagcttttt?gttgaagaga?tttgg????????????????????????????????????25

<210>53

<211>23

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(23)

<223>

<400>53

gaaaatactt?catcagcatt?acc??????????????????????????????????????23

<210>54

<211>28

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(28)

<223>

<400>54

gtcgactacg?taagtttctg?cttctacc?????????????????????????????????28

<210>55

<211>26

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(26)

<223>

<400>55

ggatccggtg?atacctgcac?atcaac???????????????????????????????????26

<210>56

<211>28

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(28)

<223>

<400>56

aagcttgcac?gaggcaaagc?aaaggttg?????????????????????????????????28

<210>57

<211>29

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(29)

<223>

<400>57

gtcgacaacc?aaatccagta?tacagttac????????????????????????????????29

<210>58

<211>30

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(30)

<223>

<400>58

aggatccaac?caaatccagt?atacagttac???????????????????????????????30

<210>59

<211>28

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(28)

<223>

<400>59

gaattcgcac?gaggcaaagc?aaaggttg????????????????????????????????28

<210>60

<211>25

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(25)

<223>

<400>60

aagctttgga?ttagcactga?ttgtc???????????????????????????????????25

<210>61

<211>29

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(29)

<223>

<400>61

gtcgacagaa?aatacttcat?cagcattac???????????????????????????????29

<210>62

<211>29

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(29)

<223>

<400>62

ggatccagaa?aatacttcat?cagcattac??????????????????????????????29

<210>63

<211>27

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(27)

<223>

<400>63

gaattctctt?tggattagca?ctgattg????????????????????????????????27

<210>64

<211>23

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(23)

<223>

<400>64

cgccttgtat?ctgtttggat?tgg????????????????????????????????????23

<210>65

<211>24

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(24)

<223>

<400>65

ctaacaatca?atgagtatga?gagc???????????????????????????????????24

<210>66

<211>26

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(26)

<223>

<400>66

agagcaaggc?cagcaggacc?acaacc??????????????????????????????????26

<210>67

<211>26

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(26)

<223>

<400>67

ccttgggagc?ttttgggata?ggctag??????????????????????????????????26

<210>68

<211>26

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(26)

<223>

<400>68

tcacgccttg?tatctgtttg?gattgg??????????????????????????????????26

<210>69

<211>15

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(15)

<223>

<400>69

gtcgagtatg?gagtt??????????????????????????????????????????????15

<210>70

<211>28

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(28)

<223>

<400>70

aagcttaccg?atagtaaaat?cgttagtt????????????????????????????????28

<210>71

<211>31

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(31)

<223>

<400>71

ctcgagctta?ccgatagtaa?aatcgttagt?t????????????????????????????31

<210>72

<211>28

<212>DNA

＜213〉composition sequence

<400>72

gtcgacaaca?acaacaaaca?acctttgc????????????????????????????????28

<210>73

<211>28

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(28)

<223>

<400>73

ggatccaaca?acaacaaaca?acctttgc????????????????????????????????28

<210>74

<211>28

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(2?8)

<223>

<400>74

gtcgactttt?tgttgaagag?atttggtg???????????????????????????????28

<210>75

<211>28

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(28)

<223>

<400>75

ctcgagactc?actgatttcc?attgcttg???????????????????????????????28

<210>76

<211>22

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(22)

<223>

<400>76

gagctctaca?aattagggtt?ac??????????????????????????????????????22

<210>77

<211>23

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(23)

<223>

<400>77

aagcttatta?tttccaaatt?ccg?????????????????????????????????????23

<210>78

<211>50

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer

<222>(1)..(50)

<223>

<400>78

aagctttgca?attcatacag?aagtgagaaa?aatgcagcta?gcagcgacag???????????50

<210>79

<211>1062

<212>DNA

＜213〉haematococcus pluvialis

<220>

<221>CDS

<222>(32)..(1021)

<223>

<400>79

aagctttgca?attcatacag?aagtgagaaa?a?atg?cag?cta?gca?gcg?aca?gta???52

Met?Gln?Leu?Ala?Ala?Thr?Val

1???????????????5

atg?ttg?gag?cag?ctt?acc?gga?agc?gct?gag?gca?ctc?aag?gag?aag?gag??100

Met?Leu?Glu?Gln?Leu?Thr?Gly?Ser?Ala?Glu?Ala?Leu?Lys?Glu?Lys?Glu

10??????????????????15??????????????????20

aag?gag?gtt?gca?ggc?agc?tct?gac?gtg?ttg?cgt?aca?tgg?gcg?acc?cag????148

Lys?Glu?Val?Ala?Gly?Ser?Ser?Asp?Val?Leu?Arg?Thr?Trp?Ala?Thr?Gln

25??????????????????30??????????????????35

tac?tcg?ctt?ccg?tca?gag?gag?tca?gac?gcg?gcc?cgc?ccg?gga?ctg?aag????196

TVr?Ser?Leu?Pro?Ser?Glu?Glu?Ser?Asp?Ala?Ala?Arg?Pro?Gly?Leu?Lys

40??????????????????45??????????????????50??????????????????55

aat?gcc?tac?aag?cca?cca?cct?tcc?gac?aca?aag?ggc?atc?aca?atg?gcg????244

Asn?Ala?Tyr?Lys?Pro?Pro?Pro?Ser?Asp?Thr?Lys?Gly?Ile?Thr?Met?Ala

60??????????????????65??????????????????70

cta?gct?gtc?atc?ggc?tcc?tgg?gcc?gca?gtg?ttc?ctc?cac?gcc?att?ttt????292

Leu?Ala?Val?Ile?Gly?Ser?Trp?Ala?Ala?Val?Phe?Leu?His?Ala?Ile?Phe

75??????????????????80??????????????????85

caa?atc?aag?ctt?ccg?acc?tcc?ttg?gac?cag?ctg?cac?tgg?ctg?ccc?gtg????340

Gln?Ile?Lys?Leu?Pro?Thr?Ser?Leu?Asp?Gln?Leu?His?Trp?Leu?Pro?Val

90??????????????????95??????????????????100

tca?gat?gcc?aca?gct?cag?ctg?gtt?agc?ggc?agc?agc?agc?ctg?ctg?cac????388

Ser?Asp?Ala?Thr?Ala?Gln?Leu?Val?Ser?Gly?Ser?Ser?Ser?Leu?Leu?His

105?????????????????110?????????????????115

atc?gtc?gta?gta?ttc?ttt?gtc?ctg?gag?ttc?ctg?tac?aca?ggc?ctt?ttt????436

Ile?Val?Val?Val?Phe?Phe?Val?Leu?Glu?Phe?Leu?Tyr?Thr?Gly?Leu?Phe

120?????????????????125?????????????????130?????????????????135

atc?acc?acg?cat?gat?gct?atg?cat?ggc?acc?atc?gcc?atg?aga?aac?agg????484

Ile?Thr?Thr?His?Asp?Ala?Met?His?Gly?Thr?Ile?Ala?Met?Arg?Asn?Arg

140?????????????????145?????????????????150

cag?ctt?aat?gac?ttc?ttg?ggc?aga?gta?tgc?atc?tcc?ttg?tac?gcc?tgg????532

Gln?Leu?Asn?Asp?Phe?Leu?Gly?Arg?Val?Cys?Ile?Ser?Leu?Tyr?Ala?Trp

155?????????????????160?????????????????165

ttt?gat?tac?aac?atg?ctg?cac?cgc?aag?cat?tgg?gag?cac?cac?aac?cac????580

Phe?Asp?Tyr?Asn?Met?Leu?His?Arg?Lys?His?Trp?Glu?His?His?Asn?His

170?????????????????175?????????????????180

act?ggc?gag?gtg?ggc?aag?gac?cct?gac?ttc?cac?agg?gga?aac?cct?ggc????628

Thr?Gly?Glu?Val?Gly?Lys?Asp?Pro?Asp?Phe?His?Arg?Gly?Asn?Pro?Gly

185?????????????????190?????????????????195

att?gtg?ccc?tgg?ttt?gcc?agc?ttc?atg?tcc?agc?tac?atg?tcg?atg?tgg????676

Ile?Val?Pro?Trp?Phe?Ala?Ser?Phe?Met?Ser?Ser?Tyr?Met?Ser?Met?Trp

200?????????????????205?????????????????210?????????????????215

cag?ttt?gcg?cgc?ctc?gca?tgg?tgg?acg?gtg?gtc?atg?cag?ctg?ctg?ggt????724

Gln?Phe?Ala?Arg?Leu?Ala?Trp?Trp?Thr?Val?Val?Met?Gln?Leu?Leu?Gly

220?????????????????225?????????????????230

gcg?cca?atg?gcg?aac?ctg?ctg?gtg?ttc?atg?gcg?gcc?gcg?ccc?atc?ctg????772

Ala?Pro?Met?Ala?Asn?Leu?Leu?Val?Phe?Met?Ala?Ala?Ala?Pro?Ile?Leu

235?????????????????240?????????????????245

tcc?gcc?ttc?cgc?ttg?ttc?tac?ttt?ggc?acg?tac?atg?ccc?cac?aag?cct????820

Ser?Ala?Phe?Arg?Leu?Phe?Tyr?Phe?Gly?Thr?Tyr?Met?Pro?His?Lys?Pro

250?????????????????255?????????????????260

gag?cct?ggc?gcc?gcg?tca?ggc?tct?tca?cca?gcc?gtc?atg?aac?tgg?tgg????868

Glu?Pro?Gly?Ala?Ala?Ser?Gly?Ser?Ser?Pro?Ala?Val?Met?Asn?Trp?Trp

265?????????????????270?????????????????275

aag?tcg?cgc?act?agc?cag?gcg?tcc?gac?ctg?gtc?agc?ttt?ctg?acc?tgc????916

Lys?Ser?Arg?Thr?Ser?Gln?Ala?Ser?Asp?Leu?Val?Ser?Phe?Leu?Thr?Cys

280?????????????????285?????????????????290?????????????????295

tac?cac?ttc?gac?ctg?cac?tgg?gag?cac?cac?cgc?tgg?ccc?ttt?gcc?ccc????964

Tyr?His?Phe?Asp?Leu?His?Trp?Glu?His?His?Arg?Trp?Pro?Phe?Ala?Pro

300?????????????????305?????????????????310

tgg?tgg?gag?ctg?ccc?aac?tgc?cgc?cgc?ctg?tct?ggc?cga?ggt?ctg?gtt????1012

Trp?Trp?Glu?Leu?Pro?Asn?Cys?Arg?Arg?Leu?Ser?Gly?Arg?Gly?Leu?Val

315?????????????????320?????????????????325

cct?gcc?tag?ctggacacac?tgcagtgggc?cctgctgcca?gctgggcatg?c??????????1062

Pro?Ala

<210>80

<211>329

<212>PRT

＜213〉haematococcus pluvialis

<400>80

Met?Gln?Leu?Ala?Ala?Thr?Val?Met?Leu?Glu?Gln?Leu?Thr?Gly?Ser?Ala

1???????????????5???????????????????10??????????????????15

Glu?Ala?Leu?Lys?Glu?Lys?Glu?Lys?Glu?Val?Ala?Gly?Ser?Ser?Asp?Val

20??????????????????25??????????????????30

Leu?Arg?Thr?Trp?Ala?Thr?Gln?Tyr?Ser?Leu?Pro?Ser?Glu?Glu?Ser?Asp

35??????????????????40??????????????????45

Ala?Ala?Arg?Pro?Gly?Leu?Lys?Asn?Ala?Tyr?Lys?Pro?Pro?Pro?Ser?Asp

50??????????????????55??????????????????60

Thr?Lys?Gly?Ile?Thr?Met?Ala?Leu?Ala?Val?Ile?Gly?Ser?Trp?Ala?Ala

65??????????????????70??????????????????75??????????????????80

Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile?Lys?Leu?Pro?Thr?Ser?Leu?Asp

85??????????????????90??????????????????95

Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Asp?Ala?Thr?Ala?Gln?Leu?Val?Ser

100?????????????????105?????????????????110

Gly?Ser?Ser?Ser?Leu?Leu?His?Ile?Val?Val?Val?Phe?Phe?Val?Leu?Glu

115?????????????????120?????????????????125

Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Thr?His?Asp?Ala?Met?His?Gly

130?????????????????135?????????????????140

Thr?Ile?Ala?Met?Arg?Asn?Arg?Gln?Leu?Asn?Asp?Phe?Leu?Gly?Arg?Val

145?????????????????150?????????????????155?????????????????160

Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp?Tyr?Asn?Met?Leu?His?Arg?Lys

165?????????????????170?????????????????175

His?Trp?Glu?His?His?Asn?His?Thr?Gly?Glu?Val?Gly?Lys?Asp?Pro?Asp

180?????????????????185?????????????????190

Phe?His?Arg?Gly?Asn?Pro?Gly?Ile?Val?Pro?Trp?Phe?Ala?Ser?Phe?Met

195?????????????????200?????????????????205

Ser?Ser?Tyr?Met?Ser?Met?Trp?Gln?Phe?Ala?Arg?Leu?Ala?Trp?Trp?Thr

210?????????????????215?????????????????220

Val?Val?Met?Gln?Leu?Leu?Gly?Ala?Pro?Met?Ala?Asn?Leu?Leu?Val?Phe

225?????????????????230?????????????????235?????????????????240

Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu?Phe?Tyr?Phe?Gly

245?????????????????250?????????????????255

Thr?Tyr?Met?Pro?His?Lys?Pro?Glu?Pro?Gly?Ala?Ala?Ser?Gly?Ser?Ser

260?????????????????265?????????????????270

Pro?Ala?Val?Met?Asn?Trp?Trp?Lys?Ser?Arg?Thr?Ser?Gln?Ala?Ser?Asp

275?????????????????280?????????????????285

Leu?Val?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?His?Trp?Glu?His

290?????????????????295?????????????????300

His?Arg?Trp?Pro?Phe?Ala?Pro?Trp?Trp?Glu?Leu?Pro?Asn?Cys?Arg?Arg

305?????????????????310?????????????????315?????????????????320

Leu?Ser?Gly?Arg?Gly?Leu?Val?Pro?Ala

325

<210>81

<211>789

<212>DNA

＜213〉some shape beads algae (Nostoc punctiforme)

<220>

<221>CDS

<222>(1)..(789)

<223>

<400>81

ttg?aat?ttt?tgt?gat?aaa?cca?gtt?agc?tat?tat?gtt?gca?ata?gag?caa????48

Leu?Asn?Phe?Cys?Asp?Lys?Pro?Val?Ser?Tyr?Tyr?Val?Ala?Ile?Glu?Gln

1???????????????5???????????????????10??????????????????15

tta?agt?gct?aaa?gaa?gat?act?gtt?tgg?ggg?ctg?gtg?att?gtc?ata?gta????96

Leu?Ser?Ala?Lys?Glu?Asp?Thr?Val?Trp?Gly?Leu?Val?Ile?Val?Ile?Val

20??????????????????25??????????????????30

att?att?agt?ctt?tgg?gta?gct?agt?ttg?gct?ttt?tta?cta?gct?att?aat????144

Ile?Ile?Ser?Leu?Trp?Val?Ala?Ser?Leu?Ala?Phe?Leu?Leu?Ala?Ile?Asn

35??????????????????40??????????????????45

tat?gcc?aaa?gtc?cca?att?tgg?ttg?ata?cct?att?gca?ata?gtt?tgg?caa????192

Tyr?Ala?Lys?Val?Pro?Ile?Trp?Leu?Ile?Pro?Ile?Ala?Ile?Val?Trp?Gln

50??????????????????55??????????????????60

atg?ttc?ctt?tat?aca?ggg?cta?ttt?att?act?gca?cat?gat?gct?atg?cat????240

Met?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Ala?His?Asp?Ala?Met?His

65??????????????????70??????????????????75??????????????????80

ggg?tca?gtt?tat?cgt?aaa?aat?ccc?aaa?att?aat?aat?ttt?atc?ggt?tca????288

Gly?Ser?Val?Tyr?Arg?Lys?Asn?Pro?Lys?Ile?Asn?Asn?Phe?Ile?Gly?Ser

85??????????????????90??????????????????95

cta?gct?gta?gcg?ctt?tac?gct?gtg?ttt?cca?tat?caa?cag?atg?tta?aag????336

Leu?Ala?Val?Ala?Leu?Tyr?Ala?Val?Phe?Pro?Tyr?Gln?Gln?Met?Leu?Lys

100?????????????????105?????????????????110

aat?cat?tgc?tta?cat?cat?cgt?cat?cct?gct?agc?gaa?gtt?gac?cca?gat????384

Asn?His?Cys?Leu?His?His?Arg?His?Pro?Ala?Ser?Glu?Val?Asp?Pro?Asp

115?????????????????120?????????????????125

ttt?cat?gat?ggt?aag?aga?aca?aac?gct?att?ttc?tgg?tat?ctc?cat?ttc????432

Phe?His?Asp?Gly?Lys?Arg?Thr?Asn?Ala?Ile?Phe?Trp?Tyr?Leu?His?Phe

130?????????????????135?????????????????140

atg?ata?gaa?tac?tcc?agt?tgg?caa?cag?tta?ata?gta?cta?act?atc?cta????480

Met?Ile?Glu?Tyr?Ser?Ser?Trp?Gln?Gln?Leu?Ile?Val?Leu?Thr?Ile?Leu

145?????????????????150?????????????????155?????????????????160

ttt?aat?tta?gct?aaa?tac?gtt?ttg?cac?atc?cat?caa?ata?aat?ctc?atc????528

Phe?Asn?Leu?Ala?Lys?Tyr?Val?Leu?His?Ile?His?Gln?Ile?Asn?Leu?Ile

165?????????????????170?????????????????175

tta?ttt?tgg?agt?att?cct?cca?att?tta?agt?tcc?att?caa?ctg?ttt?tat????576

Leu?Phe?Trp?Ser?Ile?Pro?Pro?Ile?Leu?Ser?Ser?Ile?Gln?Leu?Phe?Tyr

180?????????????????185?????????????????190

ttc?gga?aca?ttt?ttg?cct?cat?cga?gaa?ccc?aag?aaa?gga?tat?gtt?tat????624

Phe?Gly?Thr?Phe?Leu?Pro?His?Arg?Glu?Pro?Lys?Lys?Gly?Tyr?Val?Tyr

195?????????????????200?????????????????205

ccc?cat?tgc?agc?caa?aca?ata?aaa?ttg?cca?act?ttt?ttg?tca?ttt?atc????672

Pro?His?Cys?Ser?Gln?Thr?Ile?Lys?Leu?Pro?Thr?Phe?Leu?Ser?Phe?Ile

210?????????????????215?????????????????220

gct?tgc?tac?cac?ttt?ggt?tat?cat?gaa?gaa?cat?cat?gag?tat?ccc?cat????720

Ala?Cys?Tyr?His?Phe?Gly?Tyr?His?Glu?Glu?His?His?Glu?Tyr?Pro?His

225?????????????????230?????????????????235?????????????????240

gta?cct?tgg?tgg?caa?ctt?cca?tct?gta?tat?aag?cag?aga?gta?ttc?aac????768

Val?Pro?Trp?Trp?Gln?Leu?Pro?Ser?Val?Tyr?Lys?Gln?Arg?Val?Phe?Asn

245?????????????????250?????????????????255

aat?tca?gta?acc?aat?tcg?taa????????????????????????????????????????789

Asn?Ser?Val?Thr?Asn?Ser

260

<210>82

<211>262

<212>PRT

＜213〉some shape beads algae

<400>82

Leu?Asn?Phe?Cys?Asp?Lys?Pro?Val?Ser?Tyr?Tyr?Val?Ala?Ile?Glu?Gln

1???????????????5???????????????????10??????????????????15

Leu?Ser?Ala?Lys?Glu?Asp?Thr?Val?Trp?Gly?Leu?Val?Ile?Val?Ile?Val

20??????????????????25??????????????????30

Ile?Ile?Ser?Leu?Trp?Val?Ala?Ser?Leu?Ala?Phe?Leu?Leu?Ala?Ile?Asn

35??????????????????40??????????????????45

Tyr?Ala?Lys?Val?Pro?Ile?Trp?Leu?Ile?Pro?Ile?Ala?Ile?Val?Trp?Gln

50??????????????????55??????????????????60

Met?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Ala?His?Asp?Ala?Met?His

65??????????????????70??????????????????75??????????????????80

Gly?Ser?Val?Tyr?Arg?Lys?Asn?Pro?Lys?Ile?Asn?Asn?Phe?Ile?Gly?Ser

85??????????????????90??????????????????95

Leu?Ala?Val?Ala?Leu?Tyr?Ala?Val?Phe?Pro?Tyr?Gln?Gln?Met?Leu?Lys

100?????????????????105?????????????????110

Asn?His?Cys?Leu?His?His?Arg?His?Pro?Ala?Ser?Glu?Val?Asp?Pro?Asp

115?????????????????120?????????????????125

Phe?His?Asp?Gly?Lys?Arg?Thr?Asn?Ala?Ile?Phe?Trp?Tyr?Leu?His?Phe

130?????????????????135?????????????????140

Met?Ile?Glu?Tyr?Ser?Ser?Trp?Gln?Gln?Leu?Ile?Val?Leu?Thr?Ile?Leu

145?????????????????150?????????????????155?????????????????160

Phe?Asn?Leu?Ala?Lys?Tyr?Val?Leu?His?Ile?His?Gln?Ile?Asn?Leu?Ile

165?????????????????170?????????????????175

Leu?Phe?Trp?Ser?Ile?Pro?Pro?Ile?Leu?Ser?Ser?Ile?Gln?Leu?Phe?Tyr

180?????????????????185?????????????????190

Phe?Gly?Thr?Phe?Leu?Pro?His?Arg?Glu?Pro?Lys?Lys?Gly?Tyr?Val?Tyr

195?????????????????200?????????????????205

Pro?His?Cys?Ser?Gln?Thr?Ile?Lys?Leu?Pro?Thr?Phe?Leu?Ser?Phe?Ile

210?????????????????215?????????????????220

Ala?Cys?Tyr?His?Phe?Gly?Tyr?His?Glu?Glu?His?His?Glu?Tyr?Pro?His

225?????????????????230?????????????????235?????????????????240

Val?Pro?Trp?Trp?Gln?Leu?Pro?Ser?Val?Tyr?Lys?Gln?Arg?Val?Phe?Asn

245?????????????????250?????????????????255

Asn?Ser?Val?Thr?Asn?Ser

260

<210>83

<211>762

<212>DNA

＜213〉some shape beads algae

<220>

<221>CDS

<222>(1)..(762)

<223>

<400>83

gtg?atc?cag?tta?gaa?caa?cca?ctc?agt?cat?caa?gca?aaa?ctg?act?cca????48

Val?Ile?Gln?Leu?Glu?Gln?Pro?Leu?Ser?His?Gln?Ala?Lys?Leu?Thr?Pro

1???????????????5???????????????????10??????????????????15

gta?ctg?aga?agt?aaa?tct?cag?ttt?aag?ggg?ctt?ttc?att?gct?att?gtc????96

Val?Leu?Arg?Ser?Lys?Ser?Gln?Phe?Lys?Gly?Leu?Phe?Ile?Ala?Ile?Val

20??????????????????25??????????????????30

att?gtt?agc?gca?tgg?gtc?att?agc?ctg?agt?tta?tta?ctt?tcc?ctt?gac????114

Ile?Val?Ser?Ala?Trp?Val?Ile?Ser?Leu?Ser?Leu?Leu?Leu?Ser?Leu?Asp

35??????????????????40??????????????????45

atc?tca?aag?cta?aaa?ttt?tgg?atg?tta?ttg?cct?gtt?ata?cta?tgg?caa????192

Ile?Ser?Lys?Leu?Lys?Phe?Trp?Met?Leu?Leu?Pro?Val?Ile?Leu?Trp?Gln

50??????????????????55??????????????????60

aca?ttt?tta?tat?acg?gga?tta?ttt?att?aca?tct?cat?gat?gcc?atg?cat????240

Thr?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Ser?His?Asp?Ala?Met?His

65??????????????????70??????????????????75??????????????????80

ggc?gta?gta?ttt?ccc?caa?aac?acc?aag?att?aat?cat?ttg?att?gga?aca????288

Gly?Val?Val?Phe?Pro?Gln?Asn?Thr?Lys?Ile?Asn?His?Leu?Ile?Gly?Thr

85??????????????????90??????????????????95

ttg?acc?cta?tcc?ctt?tat?ggt?ctt?tta?cca?tat?caa?aaa?cta?ttg?aaa????336

Leu?Thr?Leu?Ser?Leu?Tyr?Gly?Leu?Leu?Pro?Tyr?Gln?Lys?Leu?Leu?Lys

100?????????????????105?????????????????110

aaa?cat?tgg?tta?cac?cac?cac?aat?cca?gca?agc?tca?ata?gac?ccg?gat????384

Lys?His?Trp?Leu?His?His?His?Asn?Pro?Ala?Ser?Ser?Ile?Asp?Pro?Asp

115?????????????????120?????????????????125

ttt?cac?aat?ggt?aaa?cac?caa?agt?ttc?ttt?gct?tgg?tat?ttt?cat?ttt????432

Phe?His?Asn?Gly?Lys?His?Gln?Ser?Phe?Phe?Ala?Trp?Tyr?Phe?His?Phe

130?????????????????135?????????????????140

atg?aaa?ggt?tac?tgg?agt?tgg?ggg?caa?ata?att?gcg?ttg?act?att?att????480

Met?Lys?Gly?Tyr?Trp?Ser?Trp?Gly?Gln?Ile?Ile?Ala?Leu?Thr?Ile?Ile

145?????????????????150?????????????????155?????????????????160

tat?aac?ttt?gct?aaa?tac?ata?ctc?cat?atc?cca?agt?gat?aat?cta?act????528

Tyr?Asn?Phe?Ala?Lys?Tyr?Ile?Leu?His?Ile?Pro?Ser?Asp?Asn?Leu?Thr

165?????????????????170?????????????????175

tac?ttt?tgg?gtg?cta?ccc?tcg?ctt?tta?agt?tca?tta?caa?tta?ttc?tat????576

Tyr?Phe?Trp?Val?Leu?Pro?Ser?Leu?Leu?Ser?Ser?Leu?Gln?Leu?Phe?Tyr

180?????????????????185?????????????????190

ttt?ggt?act?ttt?tta?ccc?cat?agt?gaa?cca?ata?ggg?ggt?tat?gtt?cag????624

Phe?Gly?Thr?Phe?Leu?Pro?His?Ser?Glu?Pro?Ile?Gly?Gly?Tyr?Val?Gln

195?????????????????200?????????????????205

cct?cat?tgt?gcc?caa?aca?att?agc?cgt?cct?att?tgg?tgg?tca?ttt?atc????672

Pro?His?Cys?Ala?Gln?Thr?Ile?Ser?Arg?Pro?Ile?Trp?Trp?Ser?Phe?Ile

210?????????????????215?????????????????220

acg?tgc?tat?cat?ttt?ggc?tac?cac?gag?gaa?cat?cac?gaa?tat?cct?cat????720

Thr?Cys?Tyr?His?Phe?Gly?Tyr?His?Glu?Glu?His?His?Glu?Tyr?Pro?His

225?????????????????230?????????????????235?????????????????240

att?tct?tgg?tgg?cag?tta?cca?gaa?att?tac?aaa?gca?aaa?tag????????????762

Ile?Ser?Trp?Trp?Gln?Leu?Pro?Glu?Ile?Tyr?Lys?Ala?Lys

245?????????????????250

<210>84

<211>253

<212>PRT

＜213〉some shape beads algae

<400>84

Val?Ile?Gln?Leu?Glu?Gln?Pro?Leu?Ser?His?Gln?Ala?Lys?Leu?Thr?Pro

1???????????????5???????????????????10??????????????????15

Val?Leu?Arg?Ser?Lys?Ser?Gln?Phe?Lys?Gly?Leu?Phe?Ile?Ala?Ile?Val

20??????????????????25??????????????????30

Ile?Val?Ser?Ala?Trp?Val?Ile?Ser?Leu?Ser?Leu?Leu?Leu?Ser?Leu?Asp

35??????????????????40??????????????????45

Ile?Ser?Lys?Leu?Lys?Phe?Trp?Met?Leu?Leu?Pro?Val?Ile?Leu?Trp?Gln

50??????????????????55??????????????????60

Thr?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Ser?His?Asp?Ala?Met?His

65??????????????????70??????????????????75??????????????????80

Gly?Val?Val?Phe?Pro?Gln?Asn?Thr?Lys?Ile?Asn?His?Leu?Ile?Gly?Thr

85??????????????????90??????????????????95

Leu?Thr?Leu?Ser?Leu?Tyr?Gly?Leu?Leu?Pro?Tyr?Gln?Lys?Leu?Leu?Lys

100?????????????????105?????????????????110

Lys?His?Trp?Leu?His?His?His?Asn?Pro?Ala?Ser?Ser?Ile?Asp?Pro?Asp

115?????????????????120?????????????????125

Phe?His?Asn?Gly?Lys?His?Gln?Ser?Phe?Phe?Ala?Trp?Tyr?Phe?His?Phe

130?????????????????135?????????????????140

Met?Lys?Gly?Tyr?Trp?Ser?Trp?Gly?Gln?Ile?Ile?Ala?Leu?Thr?Ile?Ile

145?????????????????150?????????????????155?????????????????160

Tyr?Asn?Phe?Ala?Lys?Tyr?Ile?Leu?His?Ile?Pro?Ser?Asp?Asn?Leu?Thr

165?????????????????170?????????????????175

Tyr?Phe?Trp?Val?Leu?Pro?Ser?Leu?Leu?Ser?Ser?Leu?Gln?Leu?Phe?Tyr

180?????????????????185?????????????????190

Phe?Gly?Thr?Phe?Leu?Pro?His?Ser?Glu?Pro?Ile?Gly?Gly?Tyr?Val?Gln

195?????????????????200?????????????????205

Pro?His?Cys?Ala?Gln?Thr?Ile?Ser?Arg?Pro?Ile?Trp?Trp?Ser?Phe?Ile

210?????????????????215?????????????????220

Thr?Cys?Tyr?His?Phe?Gly?Tyr?His?Glu?Glu?His?His?Glu?Tyr?Pro?His

225?????????????????230?????????????????235?????????????????240

Ile?Ser?Trp?Trp?Gln?Leu?Pro?Glu?Ile?Tyr?Lys?Ala?Lys

245?????????????????250

<210>85

<211>804

<212>DNA

＜213〉Synechococcus (synechococcus) WH8102

<220>

<221>CDS

<222>(1)..(804)

<223>

<400>85

atg?aaa?acg?aca?aga?tct?att?tcg?tgg?cca?tcg?act?tgc?tgg?cat?cac????48

Met?Lys?Thr?Thr?Arg?Ser?Ile?Ser?Trp?Pro?Ser?Thr?Cys?Trp?His?His

1???????????????5???????????????????10??????????????????15

cag?ccg?agt?tgc?tca?agc?tgg?gtg?gca?aat?gag?ttc?agc?cct?cag?gcc????96

Gln?Pro?Ser?Cys?Ser?Ser?Trp?Val?Ala?Asn?Glu?Phe?Ser?Pro?Gln?Ala

20??????????????????25??????????????????30

ctc?aaa?ggg?ttg?gct?ctg?gct?ggt?ctg?att?gga?tca?gcc?tgg?ctg?ctc????144

Leu?Lys?Gly?Leu?Ala?Leu?Ala?Gly?Leu?Ile?Gly?Ser?Ala?Trp?Leu?Leu

35??????????????????40??????????????????45

tcc?ctg?ggc?ctg?agc?tac?acc?ctg?cca?ctt?gat?cag?acg?cct?ggg?ctg????192

Ser?Leu?Gly?Leu?Ser?Tyr?Thr?Leu?Pro?Leu?Asp?Gln?Thr?Pro?Gly?Leu

50??????????????????55??????????????????60

ttg?att?ggc?agc?ttg?att?ctg?ctc?aga?gca?ttt?ctg?cac?acc?ggg?ctg????240

Leu?Ile?Gly?Ser?Leu?Ile?Leu?Leu?Arg?Ala?Phe?Leu?His?Thr?Gly?Leu

65??????????????????70??????????????????75??????????????????80

ttc?atc?gtt?gcc?cac?gat?tcc?atg?cac?gcc?agt?ctg?gtt?ccg?ggt?cat????288

Phe?Ile?Val?Ala?His?Asp?Ser?Met?His?Ala?Ser?Leu?Val?Pro?Gly?His

85??????????????????90??????????????????95

ccc?gga?ttg?aac?cgc?tgg?atc?ggc?aaa?gtg?tat?ttg?ttg?gtg?tat?gca????336

Pro?Gly?Leu?Asn?Arg?Trp?Ile?Gly?Lys?Val?Tyr?Leu?Leu?Val?Tyr?Ala

100?????????????????105?????????????????110

ggc?ttg?tct?tat?gag?cgt?tgt?tcc?cgc?aac?cac?aga?cgt?cat?cac?ctg????384

Gly?Leu?Ser?Tyr?Glu?Arg?Cys?Ser?Arg?Asn?His?Arg?Arg?His?His?Leu

115?????????????????120?????????????????125

gca?ccg?gag?acg?ttc?cag?gat?cct?gac?tac?caa?cgt?tgc?acc?aat?aac????432

Ala?Pro?Glu?Thr?Phe?Gln?Asp?Pro?Asp?Tyr?Gln?Arg?Cys?Thr?Asn?Asn

130?????????????????135?????????????????140

aac?atc?cta?gat?tgg?tat?gtt?cac?ttc?atg?ggc?aac?tat?ctg?ggc?atg????480

Asn?Ile?Leu?Asp?Trp?Tyr?Val?His?Phe?Met?Gly?Asn?Tyr?Leu?Gly?Met

145?????????????????150?????????????????155?????????????????160

cgg?caa?ctg?tta?aat?cta?agc?tgt?ctt?tgg?ctg?gcg?cta?atc?att?ctc????528

Arg?Gln?Leu?Leu?Asn?Leu?Ser?Cys?Leu?Trp?Leu?Ala?Leu?Ile?Ile?Leu

165?????????????????170?????????????????175

aac?ggt?tct?gat?ctc?cct?gct?cag?atc?atg?cat?ctg?ctg?ttg?ttc?agc????576

Asn?Gly?Ser?Asp?Leu?Pro?Ala?Gln?Ile?Met?His?Leu?Leu?Leu?Phe?Ser

180?????????????????185?????????????????190

gtt?ctg?ccg?ttg?atc?atc?agt?tcc?tgt?caa?ttg?ttt?cta?gtg?gga?acc????624

Val?Leu?Pro?Leu?Ile?Ile?Ser?Ser?Cys?Gln?Leu?Phe?Leu?Val?Gly?Thr

195?????????????????200?????????????????205

tgg?tta?ccc?cac?cga?cgt?ggg?gcc?acg?aca?cga?ccg?ggc?gtg?aca?acg????672

Trp?Leu?Pro?His?Arg?Arg?Gly?Ala?Thr?Thr?Arg?Pro?Gly?Val?Thr?Thr

210?????????????????215?????????????????220

cgc?agc?ctg?gct?ttg?cat?cca?gcc?ctc?tct?ttc?gca?gct?tgt?tac?aac????720

Arg?Ser?Leu?Ala?Leu?His?Pro?Ala?Leu?Ser?Phe?Ala?Ala?Cys?Tyr?Asn

225?????????????????230?????????????????235?????????????????240

ttt?ggc?tat?cat?cgt?gaa?cat?cat?gaa?tcg?cct?tcc?aca?ccc?tgg?ttt????768

Phe?Gly?Tyr?His?Arg?Glu?His?His?Glu?Ser?Pro?Ser?Thr?Pro?Trp?Phe

245?????????????????250?????????????????255

cag?ctg?cca?caa?ctt?cga?aat?gaa?tca?ttc?act?tga????????????????????804

Gln?Leu?Pro?Gln?Leu?Arg?Asn?Glu?Ser?Phe?Thr

260?????????????????265

<210>86

<211>267

<212>PRT

＜213〉Synechococcus WH8102

<400>86

Met?Lys?Thr?Thr?Arg?Ser?Ile?Ser?Trp?Pro?Ser?Thr?Cys?Trp?His?His

1???????????????5???????????????????10??????????????????15

Gln?Pro?Ser?Cys?Ser?Ser?Trp?Val?Ala?Asn?Glu?Phe?Ser?Pro?Gln?Ala

20??????????????????25??????????????????30

Leu?Lys?Gly?Leu?Ala?Leu?Ala?Gly?Leu?Ile?Gly?Ser?Ala?Trp?Leu?Leu

35??????????????????40??????????????????45

Ser?Leu?Gly?Leu?Ser?Tyr?Thr?Leu?Pro?Leu?Asp?Gln?Thr?Pro?Gly?Leu

50??????????????????55??????????????????60

Leu?Ile?Gly?Ser?Leu?Ile?Leu?Leu?Arg?Ala?Phe?Leu?His?Thr?Gly?Leu

65??????????????????70??????????????????75??????????????????80

Phe?Ile?Val?Ala?His?Asp?Ser?Met?His?Ala?Ser?Leu?Val?Pro?Gly?His

85??????????????????90??????????????????95

Pro?Gly?Leu?Asn?Arg?Trp?Ile?Gly?Lys?Val?Tyr?Leu?Leu?Val?Tyr?Ala

100?????????????????105?????????????????110

Gly?Leu?Ser?Tyr?Glu?Arg?Cys?Ser?Arg?Asn?His?Arg?Arg?His?His?Leu

115?????????????????120?????????????????125

Ala?Pro?Glu?Thr?Phe?Gln?Asp?Pro?Asp?Tyr?Gln?Arg?Cys?Thr?Asn?Asn

130?????????????????135?????????????????140

Asn?Ile?Leu?Asp?Trp?Tyr?Val?His?Phe?Met?Gly?Asn?Tyr?Leu?Gly?Met

145?????????????????150?????????????????155?????????????????160

Arg?Gln?Leu?Leu?Asn?Leu?Ser?Cys?Leu?Trp?Leu?Ala?Leu?Ile?Ile?Leu

165?????????????????170?????????????????175

Asn?Gly?Ser?Asp?Leu?Pro?Ala?Gln?Ile?Met?His?Leu?Leu?Leu?Phe?Ser

180?????????????????185?????????????????190

Val?Leu?Pro?Leu?Ile?Ile?Ser?Ser?Cys?Gln?Leu?Phe?Leu?Val?Gly?Thr

195?????????????????200?????????????????205

Trp?Leu?Pro?His?Arg?Arg?Gly?Ala?Thr?Thr?Arg?Pro?Gly?Val?Thr?Thr

210?????????????????215?????????????????220

Arg?Ser?Leu?Ala?Leu?His?Pro?Ala?Leu?Ser?Phe?Ala?Ala?Cys?Tyr?Asn

225?????????????????230?????????????????235?????????????????240

Phe?Gly?Tyr?His?Arg?Glu?His?His?Glu?Ser?Pro?Ser?Thr?Pro?Trp?Phe

245?????????????????250?????????????????255

Gln?Leu?Pro?Gln?Leu?Arg?Asn?Glu?Ser?Phe?Thr

260?????????????????265

<210>87

<211>33

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer _ combination

<222>(1)..(33)

<223>

<400>87

gcatgctcta?gaccttataa?agatattttg?tga???????????????????????????33

<210>88

<211>33

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer _ combination

<222>(1)..(33)

<223>

<400>88

gcatgcatct?agaaatggtt?cagtgtcaac?cat??????????????????????????33

<210>89

<211>805

<212>DNA

＜213〉beads algae PCC7120 strain

<220>

＜221〉variation

<222>(1)..(805)

<223>

<400>89

gcatgcatct?agaaatggtt?cagtgtcaac?catcatctct?gcattcagaa?aaactggtgt?????60

tattgtcatc?gacaatcaga?gatgataaaa?atattaataa?gggtatattt?attgcctgct????120

ttatcttatt?tttatgggca?attagtttaa?tcttattact?ctcaatagat?acatccataa????180

ttcataagag?cttattaggt?atagccatgc?tttggcagac?cttcttatat?acaggtttat????240

ttattactgc?tcatgatgcc?atgcacggcg?tagtttatcc?caaaaatccc?agaataaata????300

attttatagg?taagctcact?ctaatcttgt?atggactact?cccttataaa?gatttattga????360

aaaaacattg?gttacaccac?ggacatcctg?gtactgattt?agaccctgat?tattacaatg????420

gtcatcccca?aaacttcttt?ctttggtatc?tacattttat?gaagtcttat?tggcgatgga????480

cgcaaatttt?cggattagtg?atgatttttc?atggacttaa?aaatctggtg?catataccag????540

aaaataattt?aattatattt?tggatgatac?cttctatttt?aagttcagta?caactatttt????600

attttggtac?atttttgcct?cataaaaagc?tagaaggtgg?ttatactaac?ccccattgtg????660

cgcgcagtat?cccattacct?cttttttggt?cttttgttac?ttgttatcac?ttcggctacc????720

acaaggaaca?tcacgaatac?cctcaacttc?cttggtggaa?attacctgaa?gctcacaaaa????780

tatctttata?aggtctagag?catgc??????????????????????????????????????????805

<210>90

<211>35

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer _ combination

<222>(1)..(35)

<223>

<400>90

gagctcttca?ttatttcgat?tttgatttcg?tgacc???????????????????????????????35

<210>91

<211>44

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer _ combination

<222>(1)..(44)

<223>

<400>91

aagcttgagc?tcggttgatc?agaagaagaa?gaagaagatg?aact???????????????44

<210>92

<211>653

<212>DNA

＜213〉mouseearcress

<220>

＜221〉promoter

<222>(1)..(653)

<223>

<400>92

gagctcttca?ttatttcgat?tttgatttcg?tgaccagcga?acgcagaata?ccttgttgtg?????60

taatacttta?cccgtgtaaa?tcaaaaacaa?aaaggctttt?gagctttttg?tagttgaatt????120

tctctggctg?atcttttctg?tacagattca?tatatctgca?gagacgatat?cattgattat????180

ttgagcttct?tttgaactat?ttcgtgtaat?ttgggatgag?agctctatgt?atgtgtgtaa????240

actttgaaga?caacaagaaa?ggtaacaagt?gagggaggga?tgactccatg?tcaaaataga????300

tgtcataaga?ggcccatcaa?taagtgcttg?agcccattag?ctagcccagt?aactaccaga????360

ttgtgagatg?gatgtgtgaa?cagttttttt?tttgatgtag?gactgaaatg?tgaacaacag????420

gcgcatgaaa?ggctaaatta?ggacaatgat?aagcagaaat?aacttatcct?ctctaacact????480

tggcctcaca?ttgcccttca?cacaatccac?acacatccaa?tcacaacctc?atcatatatc????540

tcccgctaat?ctttttttct?ttgatctttt?tttttttgct?tattattttt?ttgactttga????600

tctcccatca?gttcatcttc?ttcttcttct?tctgatcaac?cgagctcaag?ctt???????????653

<210>93

<211>28

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer _ combination

<222>(1)..(28)

<223>

<400>93

gagctcactc?actgatttcc?attgcttg???????????????????????????????????????28

<210>94

<211>30

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer _ combination

<222>(1)..(30)

<223>

<400>94

aagcttgagc?tctttgttga?agagatttgg????????????????????????????????30

<210>95

<211>37

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer _ combination

<222>(1)..(37)

<223>

<400>95

cgccgttaag?tcgatgtccg?ttgatttaaa?cagtgtc????????????????????????37

<210>96

<211>34

<212>DNA

＜213〉composition sequence

<220>

＜221〉primer _ combination

<222>(1)..(34)

<223>

<400>96

atcaacggac?atcgacttaa?cggcgtttgt?aaac?????????????????????????????34

<210>97

<211>831

<212>DNA

＜213〉haematococcus pluvialis

<220>

<221>CDS

<222>(1)..(831)

<223>

<400>97

atg?cca?tcc?gag?tcg?tca?gac?gca?gct?cgt?cct?gtg?ttg?aag?cac?gcc????48

Met?Pro?Ser?Glu?Ser?Ser?Asp?Ala?Ala?Arg?Pro?Val?Leu?Lys?His?Ala

1???????????????5???????????????????10??????????????????15

tat?aaa?cct?cca?gca?tct?gac?gcc?aag?ggc?atc?act?atg?gcg?ctg?acc????96

Tyr?Lys?Pro?Pro?Ala?Ser?Asp?Ala?Lys?Gly?Ile?Thr?Met?Ala?Leu?Thr

20??????????????????25??????????????????30

atc?att?ggc?acc?tgg?acc?gca?gtg?ttt?tta?cac?gca?ata?ttc?caa?atc????144

Ile?Ile?Gly?Thr?Trp?Thr?Ala?Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile

35??????????????????40??????????????????45

agg?cta?ccg?aca?tcc?atg?gac?cag?ctt?cac?tgg?ttg?cct?gtg?tcc?gaa????192

Arg?Leu?Pro?Thr?Ser?Met?Asp?Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Glu

50??????????????????55??????????????????60

gcc?aca?gcc?cag?ctg?ttg?ggc?gga?agc?agc?agc?cta?ttg?cac?atc?gcc????240

Ala?Thr?Ala?Gln?Leu?Leu?Gly?Gly?Ser?Ser?Ser?Leu?Leu?His?Ile?Ala

65??????????????????70??????????????????75??????????????????80

gca?gtc?ttc?att?gta?ctt?gag?ttt?ctg?tac?act?ggt?cta?ttc?atc?acc????288

Ala?Val?Phe?Ile?Val?Leu?Glu?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr

85??????????????????90??????????????????95

acg?cat?gat?gca?atg?cat?ggc?acc?ata?gct?ttg?agg?aac?agg?cag?ctc????336

Thr?His?Asp?Ala?Met?His?Gly?Thr?Ile?Ala?Leu?Arg?Asn?Arg?Gln?Leu

100?????????????????105?????????????????110

aat?gat?ctc?ctt?ggc?aac?atc?tgc?ata?tca?ctg?tac?gcc?tgg?ttt?gac????384

Asn?Asp?Leu?Leu?Gly?Asn?Ile?Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp

115?????????????????120?????????????????125

tac?agc?atg?cac?tgg?gag?cac?cac?aac?cat?act?ggc?gaa?gtg?ggg?aaa????432

Tyr?Ser?Met?His?Trp?Glu?His?His?Asn?His?Thr?Gly?Glu?Val?Gly?Lys

130?????????????????135?????????????????140

gac?cct?gac?ttc?cac?aaa?gga?aat?cct?ggc?ctt?gtc?ccc?tgg?ttc?gcc????480

Asp?Pro?Asp?Phe?His?Lys?Gly?Asn?Pro?Gly?Leu?Val?Pro?Trp?Phe?Ala

145?????????????????150?????????????????155?????????????????160

agc?ttc?atg?tcc?agc?tac?atg?tcc?ctg?tgg?cag?ttt?gcc?cgg?ctg?gca????528

Ser?Phe?Met?Ser?Ser?Tyr?Met?Ser?Leu?Trp?Gln?Phe?Ala?Arg?Leu?Ala

165?????????????????170?????????????????155

tgg?tgg?gca?gtg?gtg?atg?caa?acg?ttg?ggg?gcc?ccc?atg?gcg?aat?ctc????576

Trp?Trp?Ala?Val?Val?Met?Gln?Thr?Leu?Gly?Ala?Pro?Met?Ala?Asn?Leu

180?????????????????185?????????????????190

cta?gtc?ttc?atg?gct?gca?gcc?cca?atc?ttg?tca?gca?ttc?cgc?ctc?ttc????624

Leu?Val?Phe?Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu?Phe

195?????????????????200?????????????????205

tac?ttc?ggc?act?tac?ctg?cca?cac?aag?cct?gag?cca?ggc?cct?gca?gca????672

Tyr?Phe?Gly?Thr?Tyr?Leu?Pro?His?Lys?Pro?Glu?Pro?Gly?Pro?Ala?Ala

210?????????????????215?????????????????220

ggc?tct?cag?gtc?atg?tct?tgg?ttc?agg?gcc?aag?aca?agt?gag?gca?tct????720

Gly?Ser?Gln?Val?Met?Ser?Trp?Phe?Arg?Ala?Lys?Thr?Ser?Glu?Ala?Ser

225?????????????????230?????????????????235?????????????????240

gat?gtg?atg?agc?ttc?ctg?aca?tgc?tac?cac?ttt?gac?ctg?ttt?gcc?ccc????768

Asp?Val?Met?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?Phe?Ala?Pro

245?????????????????250?????????????????255

tgg?tgg?cag?ctg?ccc?cac?tgc?cgc?cgc?ctg?tct?ggg?cgt?ggc?ctg?gtg????816

Trp?Trp?Gln?Leu?Pro?His?Cys?Arg?Arg?Leu?Ser?Gly?Arg?Gly?Leu?Val

260?????????????????265?????????????????270

cct?gcc?ttg?gca?tga????????????????????????????????????????????????831

Pro?Ala?Leu?Ala

275

<210>98

<211>276

<212>PRT

＜213〉haematococcus pluvialis

<400>98

Met?Pro?Ser?Glu?Ser?Ser?Asp?Ala?Ala?Arg?Pro?Val?Leu?Lys?His?Ala

1???????????????5???????????????????10??????????????????15

Tyr?Lys?Pro?Pro?Ala?Ser?Asp?Ala?Lys?Gly?Ile?Thr?Met?Ala?Leu?Thr

20??????????????????25??????????????????30

Ile?Ile?Gly?Thr?Trp?Thr?Ala?Val?Phe?Leu?His?Ala?Ile?Phe?Gln?Ile

35??????????????????40??????????????????45

Arg?Leu?Pro?Thr?Ser?Met?Asp?Gln?Leu?His?Trp?Leu?Pro?Val?Ser?Glu

50??????????????????55??????????????????60

Ala?Thr?Ala?Gln?Leu?Leu?Gly?Gly?Ser?Ser?Ser?Leu?Leu?His?Ile?Ala

65??????????????????70??????????????????75??????????????????80

Ala?Val?Phe?Ile?Val?Leu?Glu?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr

85??????????????????90??????????????????95

Thr?His?Asp?Ala?Met?His?Gly?Thr?Ile?Ala?Leu?Arg?Asn?Arg?Gln?Leu

100?????????????????105?????????????????110

Asn?Asp?Leu?Leu?Gly?Asn?Ile?Cys?Ile?Ser?Leu?Tyr?Ala?Trp?Phe?Asp

115?????????????????120?????????????????125

Tyr?Ser?Met?His?Trp?Glu?His?His?Asn?His?Thr?Gly?Glu?Val?Gly?Lys

130?????????????????135?????????????????140

Asp?Pro?Asp?Phe?His?Lys?Gly?Asn?Pro?Gly?Leu?Val?Pro?Trp?Phe?Ala

145?????????????????150?????????????????155?????????????????160

Ser?Phe?Met?Ser?Ser?Tyr?Met?Ser?Leu?Trp?Gln?Phe?Ala?Arg?Leu?Ala

165?????????????????170?????????????????175

Trp?Trp?Ala?Val?Val?Met?Gln?Thr?Leu?Gly?Ala?Pro?Met?Ala?Asn?Leu

180?????????????????185?????????????????190

Leu?Val?Phe?Met?Ala?Ala?Ala?Pro?Ile?Leu?Ser?Ala?Phe?Arg?Leu?Phe

195?????????????????200?????????????????205

Tyr?Phe?Gly?Thr?Tyr?Leu?Pro?His?Lys?Pro?Glu?Pro?Gly?Pro?Ala?Ala

210?????????????????215?????????????????220

Gly?Ser?Gln?Val?Met?Ser?Trp?Phe?Arg?Ala?Lys?Thr?Ser?Glu?Ala?Ser

225?????????????????230?????????????????235?????????????????240

Asp?Val?Met?Ser?Phe?Leu?Thr?Cys?Tyr?His?Phe?Asp?Leu?Phe?Ala?Pro

245?????????????????250?????????????????255

Trp?Trp?Gln?Leu?Pro?His?Cys?Arg?Arg?Leu?Ser?Gly?Arg?Gly?Leu?Val

260?????????????????265?????????????????270

Pro?Ala?Leu?Ala

275

<210>99

<211>729

<212>DNA

＜213〉secondary coccus (Paracoccus sp.) MBIC1143

<220>

<221>CDS

<222>(1)..(729)

<223>

<400>99

atg?agc?gca?cat?gcc?ctg?ccc?aag?gca?gat?ctg?acc?gcc?acc?agc?ctg????48

Met?Ser?Ala?His?Ala?Leu?Pro?Lys?Ala?Asp?Leu?Thr?Ala?Thr?Ser?Leu

1???????????????5???????????????????10??????????????????15

atc?gtc?tcg?ggc?ggc?atc?atc?gcc?gct?tgg?ctg?gcc?ctg?cat?gtg?cat????96

Ile?Val?Ser?Gly?Gly?Ile?Ile?Ala?Ala?Trp?Leu?Ala?Leu?His?Val?His

20??????????????????25??????????????????30

gcg?ctg?tgg?ttt?ctg?gac?gca?gcg?gcg?cat?ccc?atc?ctg?gcg?atc?gca????144

Ala?Leu?Trp?Phe?Leu?Asp?Ala?Ala?Ala?His?Pro?Ile?Leu?Ala?Ile?Ala

35??????????????????40??????????????????45

aat?ttc?ctg?ggg?ctg?acc?tgg?ctg?tcg?gtc?gga?ttg?ttc?atc?atc?gcg????192

Asn?Phe?Leu?Gly?Leu?Thr?Trp?Leu?Ser?Val?Gly?Leu?Phe?Ile?Ile?Ala

50??????????????????55??????????????????60

cat?gac?gcg?atg?cac?ggg?tcg?gtg?gtg?ccg?ggg?cgt?ccg?cgc?gcc?aat????240

His?Asp?Ala?Met?His?Gly?Ser?Val?Val?Pro?Gly?Arg?Pro?Arg?Ala?Asn

65??????????????????70??????????????????75??????????????????80

gcg?gcg?atg?ggc?cag?ctt?gtc?ctg?tgg?ctg?tat?gcc?gga?ttt?tcg?tgg????288

Ala?Ala?Met?Gly?Gln?Leu?Val?Leu?Trp?Leu?Tyr?Ala?Gly?Phe?Ser?Trp

85??????????????????90??????????????????95

cgc?aag?atg?atc?gtc?aag?cac?atg?gcc?cat?cac?cgc?cat?gcc?gga?acc????336

Arg?Lys?Met?Ile?Val?Lys?His?Met?Ala?His?His?Arg?His?Ala?Gly?Thr

100?????????????????105?????????????????110

gac?gac?gac?ccc?gat?ttc?gac?cat?ggc?ggc?ccg?gtc?cgc?tgg?tac?gcc????384

Asp?Asp?Asp?Pro?Asp?Phe?Asp?His?Gly?Gly?Pro?Val?Arg?Trp?Tyr?Ala

115?????????????????120?????????????????125

cgc?ttc?atc?ggc?acc?tat?ttc?ggc?tgg?cgc?gag?ggg?ctg?ctg?ctg?ccc????432

Arg?Phe?Ile?Gly?Thr?Tyr?Phe?Gly?Trp?Arg?Glu?Gly?Leu?Leu?Leu?Pro

130?????????????????135?????????????????140

gtc?atc?gtg?acg?gtc?tat?gcg?ctg?atc?ctt?ggg?gat?cgc?tgg?atg?tac????480

Val?Ile?Val?Thr?Val?Tyr?Ala?Leu?Ile?Leu?Gly?Asp?Arg?Trp?Met?Tyr

145?????????????????150?????????????????155?????????????????160

gtg?gtc?ttc?tgg?ccg?ctg?ccg?tcg?atc?ctg?gcg?tcg?atc?cag?ctg?ttc????528

Val?Val?Phe?Trp?Pro?Leu?Pro?Ser?Ile?Leu?Ala?Ser?Ile?Gln?Leu?Phe

165?????????????????170?????????????????175

gtg?ttc?ggc?acc?tgg?ctg?ccg?cac?cgc?ccc?ggc?cac?gac?gcg?ttc?ccg????576

Val?Phe?Gly?Thr?Trp?Leu?Pro?His?Arg?Pro?Gly?His?Asp?Ala?Phe?Pro

180?????????????????185?????????????????190

gac?cgc?cac?aat?gcg?cgg?tcg?tcg?cgg?atc?agc?gac?ccc?gtg?tcg?ctg????624

Asp?Arg?His?Asn?Ala?Arg?Ser?Ser?Arg?Ile?Ser?Asp?Pro?Val?Ser?Leu

195?????????????????200?????????????????205

ctg?acc?tgc?ttt?cac?ttt?ggc?ggt?tat?cat?cac?gaa?cac?cac?ctg?cac????672

Leu?Thr?Cys?Phe?His?Phe?Gly?Gly?Tyr?His?His?Glu?His?His?Leu?His

210?????????????????215?????????????????220

ccg?acg?gtg?ccg?tgg?tgg?cgc?ctg?ccc?agc?acc?cgc?acc?aag?ggg?gac????720

Pro?Thr?Val?Pro?Trp?Trp?Arg?Leu?Pro?Ser?Thr?Arg?Thr?Lys?Gly?Asp

225?????????????????230?????????????????235?????????????????240

acc?gca?tga????????????????????????????????????????????????????????729

Thr?Ala

<210>100

<211>242

<212>PRT

＜213〉secondary coccus MBIC1143

<400>100

Met?Ser?Ala?His?Ala?Leu?Pro?Lys?Ala?Asp?Leu?Thr?Ala?Thr?Ser?Leu

1???????????????5???????????????????10??????????????????15

Ile?Val?Ser?Gly?Gly?Ile?Ile?Ala?Ala?Trp?Leu?Ala?Leu?His?Val?His

20??????????????????25??????????????????30

Ala?Leu?Trp?Phe?Leu?Asp?Ala?Ala?Ala?His?Pro?Ile?Leu?Ala?Ile?Ala

35??????????????????40??????????????????45

Asn?Phe?Leu?Gly?Leu?Thr?Trp?Leu?Ser?Val?Gly?Leu?Phe?Ile?Ile?Ala

50??????????????????55??????????????????60

His?Asp?Ala?Met?His?Gly?Ser?Val?Val?Pro?Gly?Arg?Pro?Arg?Ala?Asn

65??????????????????70??????????????????75??????????????????80

Ala?Ala?Met?Gly?Gln?Leu?Val?Leu?Trp?Leu?Tyr?Ala?Gly?Phe?Ser?Trp

85??????????????????90??????????????????95

Arg?Lys?Met?Ile?Val?Lys?His?Met?Ala?His?His?Arg?His?Ala?Gly?Thr

100?????????????????105?????????????????110

Asp?Asp?Asp?Pro?Asp?Phe?Asp?His?Gly?Gly?Pro?Val?Arg?Trp?Tyr?Ala

115?????????????????120?????????????????125

Arg?Phe?Ile?Gly?Thr?Tyr?Phe?Gly?Trp?Arg?Glu?Gly?Leu?Leu?Leu?Pro

130?????????????????135?????????????????140

Val?Ile?Val?Thr?Val?Tyr?Ala?Leu?Ile?Leu?Gly?Asp?Arg?Trp?Met?Tyr

145?????????????????150?????????????????155?????????????????160

Val?Val?Phe?Trp?Pro?Leu?Pro?Ser?Ile?Leu?Ala?Ser?Ile?Gln?Leu?Phe

165?????????????????170?????????????????175

Val?Phe?Gly?Thr?Trp?Leu?Pro?His?Arg?Pro?Gly?His?Asp?Ala?Phe?Pro

180?????????????????185?????????????????190

Asp?Arg?His?Asn?Ala?Arg?Ser?Ser?Arg?Ile?Ser?Asp?Pro?Val?Ser?Leu

195?????????????????200?????????????????205

Leu?Thr?Cys?Phe?His?Phe?Gly?Gly?Tyr?His?His?Glu?His?His?Leu?His

210?????????????????215?????????????????220

Pro?Thr?Val?Pro?Trp?Trp?Arg?Leu?Pro?Ser?Thr?Arg?Thr?Lys?Gly?Asp

225?????????????????230?????????????????235?????????????????240

Thr?Ala

<210>101

<211>735

<212>DNA

＜213〉orange shortwave monad (Brevundimonas aurantiaca)

<220>

<221>CDS

<222>(1)..(735)

<223>

<400>101

atg?acc?gcc?gcc?gtc?gcc?gag?cca?cgc?acc?gtc?ccg?cgc?cag?acc?tgg????48

Met?Thr?Ala?Ala?Val?Ala?Glu?Pro?Arg?Thr?Val?Pro?Arg?Gln?Thr?Trp

1???????????????5???????????????????10??????????????????15

atc?ggt?ctg?acc?ctg?gcg?gga?atg?atc?gtg?gcg?gga?tgg?gcg?gtt?ctg????96

Ile?Gly?Leu?Thr?Leu?Ala?Gly?Met?Ile?Val?Ala?Gly?Trp?Ala?Val?Leu

20??????????????????25??????????????????30

cat?gtc?tac?ggc?gtc?tat?ttt?cac?cga?tgg?ggg?ccg?ttg?acc?ctg?gtg????144

His?Val?Tyr?Gly?Val?Tyr?Phe?His?Arg?Trp?Gly?Pro?Leu?Thr?Leu?Val

35??????????????????40??????????????????45

atc?gcc?ccg?gcg?atc?gtg?gcg?gtc?cag?acc?tgg?ttg?tcg?gtc?ggc?ctt????192

Ile?Ala?Pro?Ala?Ile?Val?Ala?Val?Gln?Thr?Trp?Leu?Ser?Val?Gly?Leu

50??????????????????55??????????????????60

ttc?atc?gtc?gcc?cat?gac?gcc?atg?tac?ggc?tcc?ctg?gcg?ccg?gga?cgg????240

Phe?Ile?Val?Ala?His?Asp?Ala?Met?Tyr?Gly?Ser?Leu?Ala?Pro?Gly?Arg

65??????????????????70??????????????????75??????????????????80

ccg?cgg?ctg?aac?gcc?gca?gtc?ggc?cgg?ctg?acc?ctg?ggg?ctc?tat?gcg????288

Pro?Arg?Leu?Asn?Ala?Ala?Val?Gly?Arg?Leu?Thr?Leu?Gly?Leu?Tyr?Ala

85??????????????????90??????????????????????95

ggc?ttc?cgc?ttc?gat?cgg?ctg?aag?acg?gcg?cac?cac?gcc?cac?cac?gcc????336

Gly?Phe?Arg?Phe?Asp?Arg?Leu?Lys?Thr?Ala?His?His?Ala?His?His?Ala

100?????????????????105?????????????????????110

gcg?ccc?ggc?acg?gcc?gac?gac?ccg?gat?ttt?cac?gcc?ccg?gcg?ccc?cgc????384

Ala?Pro?Gly?Thr?Ala?Asp?Asp?Pro?Asp?Phe?His?Ala?Pro?Ala?Pro?Arg

115?????????????????120?????????????????????125

gcc?ttc?ctt?ccc?tgg?ttc?ctg?aac?ttc?ttt?cgc?acc?tat?ttc?ggc?tgg????432

Ala?Phe?Leu?Pro?Trp?Phe?Leu?Asn?Phe?Phe?Arg?Thr?Tyr?Phe?Gly?Trp

130?????????????????135?????????????????????140

cgc?gag?atg?gcg?gtc?ctg?acc?gcc?ctg?gtc?ctg?atc?gcc?ctc?ttc?ggc????480

Arg?Glu?Met?Ala?Val?Leu?Thr?Ala?Leu?Val?Leu?Ile?Ala?Leu?Phe?Gly

145?????????????????150?????????????????????155?????????????160

ctg?ggg?gcg?cgg?ccg?gcc?aat?ctc?ctg?acc?ttc?tgg?gcc?gcg?ccg?gcc????528

Leu?Gly?Ala?Arg?Pro?Ala?Asn?Leu?Leu?Thr?Phe?Trp?Ala?Ala?Pro?Ala

165?????????????????????170?????????????175

ctg?ctt?tca?gcg?ctt?cag?ctc?ttc?acc?ttc?ggc?acc?tgg?ctg?ccg?cac???576

Leu?Leu?Ser?Ala?Leu?Gln?Leu?Phe?Thr?Phe?Gly?Thr?Trp?Leu?Pro?His

180?????????????????????185?????????????190

cgc?cac?acc?gac?cag?ccg?ttc?gcc?gac?gcg?cac?cac?gcc?cgc?agc?agc???624

Arg?His?Thr?Asp?Gln?Pro?Phe?Ala?Asp?Ala?His?His?Ala?Arg?Ser?Ser

195?????????????????200?????????????????205

ggc?tac?ggc?ccc?gtg?ctt?tcc?ctg?ctc?acc?tgt?ttc?cac?ttc?ggc?cgc????672

Gly?Tyr?Gly?Pro?Val?Leu?Ser?Leu?Leu?Thr?Cys?Phe?His?Phe?Gly?Arg

210?????????????????215?????????????????220

cac?cac?gaa?cac?cat?ctg?agc?ccc?tgg?cgg?ccc?tgg?tgg?cgt?ctg?tgg????720

His?His?Glu?His?His?Leu?Ser?Pro?Trp?Arg?Pro?Trp?Trp?Arg?Leu?Trp

225?????????????????230?????????????????235?????????????????240

cgc?ggc?gag?tct?tga????????????????????????????????????????????????735

Arg?Gly?Glu?Ser

<210>102

<211>244

<212>PRT

＜213〉orange shortwave monad

<400>102

Met?Thr?Ala?Ala?Val?Ala?Glu?Pro?Arg?Thr?Val?Pro?Arg?Gln?Thr?Trp

1???????????????5???????????????????10??????????????????15

Ile?Gly?Leu?Thr?Leu?Ala?Gly?Met?Ile?Val?Ala?Gly?Trp?Ala?Val?Leu

20??????????????????25??????????????????30

His?Val?Tyr?Gly?Val?Tyr?Phe?His?Arg?Trp?Gly?Pro?Leu?Thr?Leu?Val

35??????????????????40??????????????????45

Ile?Ala?Pro?Ala?Ile?Val?Ala?Val?Gln?Thr?Trp?Leu?Ser?Val?Gly?Leu

50??????????????????55??????????????????60

Phe?Ile?Val?Ala?His?Asp?Ala?Met?Tyr?Gly?Ser?Leu?Ala?Pro?Gly?Arg

65??????????????????70??????????????????75??????????????????80

Pro?Arg?Leu?Asn?Ala?Ala?Val?Gly?Arg?Leu?Thr?Leu?Gly?Leu?Tyr?Ala

85??????????????????90??????????????????95

Gly?Phe?Arg?Phe?Asp?Arg?Leu?Lys?Thr?Ala?His?His?Ala?His?His?Ala

100?????????????????105?????????????????110

Ala?Pro?Gly?Thr?Ala?Asp?Asp?Pro?Asp?Phe?His?Ala?Pro?Ala?Pro?Arg

115?????????????????120?????????????????125

Ala?Phe?Leu?Pro?Trp?Phe?Leu?Asn?Phe?Phe?Arg?Thr?Tyr?Phe?Gly?Trp

130?????????????????135?????????????????140

Arg?Glu?Met?Ala?Val?Leu?Thr?Ala?Leu?Val?Leu?Ile?Ala?Leu?Phe?Gly

145?????????????????150?????????????????155?????????????????160

Leu?Gly?Ala?Arg?Pro?Ala?Asn?Leu?Leu?Thr?Phe?Trp?Ala?Ala?Pro?Ala

165?????????????????170?????????????????175

Leu?Leu?Ser?Ala?Leu?Gln?Leu?Phe?Thr?Phe?Gly?Thr?Trp?Leu?Pro?His

180?????????????????185?????????????????190

Arg?His?Thr?Asp?Gln?Pro?Phe?Ala?Asp?Ala?His?His?Ala?Arg?Ser?Ser

195?????????????????200?????????????????205

Gly?Tyr?Gly?Pro?Val?Leu?Ser?Leu?Leu?Thr?Cys?Phe?His?Phe?Gly?Arg

210?????????????????215?????????????????220

His?His?Glu?His?His?Leu?Ser?Pro?Trp?Arg?Pro?Trp?Trp?Arg?Leu?Trp

225?????????????????230?????????????????235?????????????????240

Arg?Gly?Glu?Ser

<210>103

<211>690

<212>DNA

＜213〉foam joint ball algae (Nodularia spumigena) NSOR10

<220>

<221>CDS

<222>(1)..(690)

<223>

<400>103

atg?gcg?atc?gcc?att?att?agt?ata?tgg?gct?atc?agc?cta?ggt?ttg?tta????48

Met?Ala?Ile?Ala?Ile?Ile?Ser?Ile?Trp?Ala?Ile?Ser?Leu?Gly?Leu?Leu

1???????????????5???????????????????10??????????????????15

ctt?tat?att?gat?ata?tcc?caa?ttc?aag?ttt?tgg?atg?ttg?tta?ccg?ctc????96

Leu?Tyr?Ile?Asp?Ile?Ser?Gln?Phe?Lys?Phe?Trp?Met?Leu?Leu?Pro?Leu

20??????????????????25??????????????????30

ata?ttt?tgg?caa?aca?ttt?tta?tat?acg?gga?tta?ttt?att?aca?gct?cat????144

Ile?Phe?Trp?Gln?Thr?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Ala?His

35??????????????????40??????????????????45

gat?gcc?atg?cat?ggg?gta?gtt?ttt?ccc?aaa?aat?ccc?aaa?atc?aac?cat????192

Asp?Ala?Met?His?Gly?Val?Val?Phe?Pro?Lys?Asn?Pro?Lys?Ile?Asn?His

50??????????????????55??????????????????60

ttc?att?ggc?tca?ttg?tgc?ctg?ttt?ctt?tat?ggt?ctt?tta?cct?tat?caa????240

Phe?Ile?Gly?Ser?Leu?Cys?Leu?Phe?Leu?Tyr?Gly?Leu?Leu?Pro?Tyr?Gln

65??????????????????70??????????????????75??????????????????80

aaa?ctt?tta?aaa?aag?cat?tgg?cta?cat?cac?cat?aat?cca?gcc?agt?gaa????288

Lys?Leu?Leu?Lys?Lys?His?Trp?Leu?His?His?His?Asn?Pro?Ala?Ser?Glu

85??????????????????90??????????????????95

aca?gat?cca?gat?ttt?cac?aac?ggg?aag?cag?aaa?aac?ttt?ttt?gct?tgg????336

Thr?Asp?Pro?Asp?Phe?His?Asn?Gly?Lys?Gln?Lys?Asn?Phe?Phe?Ala?Trp

100?????????????????105?????????????????110

tat?tta?tat?ttt?atg?aag?cgt?tac?tgg?agt?tgg?tta?caa?att?atc?aca????384

Tyr?Leu?Tyr?Phe?Met?Lys?Arg?Tyr?Trp?Ser?Trp?Leu?Gln?Ile?Ile?Thr

115?????????????????120?????????????????125

tta?atg?att?att?tat?aac?tta?cta?aaa?tat?ata?tgg?cat?ttt?cca?gag????432

Leu?Met?Ile?Ile?Tyr?Asn?Leu?Leu?Lys?Tyr?Ile?Trp?His?Phe?Pro?Glu

130?????????????????135?????????????????140

gat?aat?atg?act?tat?ttt?tgg?gta?gtt?ccc?tca?att?tta?agt?tct?tta????480

Asp?Asn?Met?Thr?Tyr?Phe?Trp?Val?Val?Pro?Ser?Ile?Leu?Ser?Ser?Leu

145?????????????????150?????????????????155?????????????????160

caa?tta?ttt?tat?ttt?gga?act?ttt?cta?ccc?cac?agt?gag?cct?gta?gaa????528

Gln?Leu?Phe?Tyr?Phe?Gly?Thr?Phe?Leu?Pro?His?Ser?Glu?Pro?Val?Glu

165?????????????????170?????????????????175

ggt?tat?aaa?gag?cct?cat?cgt?tcc?caa?act?att?agc?cgt?ccc?att?tgg????576

Gly?Tyr?Lys?Glu?Pro?His?Arg?Ser?Gln?Thr?Ile?Ser?Arg?Pro?Ile?Trp

180?????????????????185?????????????????190

tgg?tca?ttt?ata?act?tgt?tac?cat?ttt?ggt?tat?cat?tac?gaa?cat?cat????624

Trp?Ser?Phe?Ile?Thr?Cys?Tyr?His?Phe?Gly?Tyr?His?Tyr?Glu?His?His

195?????????????????200?????????????????205

gaa?tac?ccc?cat?gtt?cct?tgg?tgg?caa?tta?cca?gaa?att?tat?aaa?atg????672

Glu?Tyr?Pro?His?Val?Pro?Trp?Trp?Gln?Leu?Pro?Glu?Ile?Tyr?Lys?Met

210?????????????????215?????????????????220

tct?aaa?tca?aat?ttg?tga????????????????????????????????????????????690

Ser?Lys?Ser?Asn?Leu

225

<210>104

<211>229

<212>PRT

＜213〉foam joint ball algae NSOR10

<400>104

Met?Ala?Ile?Ala?Ile?Ile?Ser?Ile?Trp?Ala?Ile?Ser?Leu?Gly?Leu?Leu

1???????????????5???????????????????10??????????????????15

Leu?Tyr?Ile?Asp?Ile?Ser?Gln?Phe?Lys?Phe?Trp?Met?Leu?Leu?Pro?Leu

20??????????????????25??????????????????30

Ile?Phe?Trp?Gln?Thr?Phe?Leu?Tyr?Thr?Gly?Leu?Phe?Ile?Thr?Ala?His

35??????????????????40??????????????????45

Asp?Ala?Met?His?Gly?Val?Val?Phe?Pro?Lys?Asn?Pro?Lys?Ile?Asn?His

50??????????????????55??????????????????60

Phe?Ile?Gly?Ser?Leu?Cys?Leu?Phe?Leu?Tyr?Gly?Leu?Leu?Pro?Tyr?Gln

65??????????????????70??????????????????75??????????????????80

Lys?Leu?Leu?Lys?Lys?His?Trp?Leu?His?His?His?Asn?Pro?Ala?Ser?Glu

85??????????????????90??????????????????95

Thr?Asp?Pro?Asp?Phe?His?Asn?Gly?Lys?Gln?Lys?Asn?Phe?Phe?Ala?Trp

100?????????????????105?????????????????110

Tyr?Leu?Tyr?Phe?Met?Lys?Arg?Tyr?Trp?Ser?Trp?Leu?Gln?Ile?Ile?Thr

115?????????????????120?????????????????125

Leu?Met?Ile?Ile?Tyr?Asn?Leu?Leu?Lys?Tyr?Ile?Trp?His?Phe?Pro?Glu

130?????????????????135?????????????????140

Asp?Asn?Met?Thr?Tyr?Phe?Trp?Val?Val?Pro?Ser?Ile?Leu?Ser?Ser?Leu

145?????????????????150?????????????????155?????????????????160

Gln?Leu?Phe?Tyr?Phe?Gly?Thr?Phe?Leu?Pro?His?Ser?Glu?Pro?Val?Glu

165?????????????????170?????????????????175

Gly?Tyr?Lys?Glu?Pro?His?Arg?Ser?Gln?Thr?Ile?Ser?Arg?Pro?Ile?Trp

180?????????????????185?????????????????190

Trp?Ser?Phe?Ile?Thr?Cys?Tyr?His?Phe?Gly?Tyr?His?Tyr?Glu?His?His

195?????????????????200?????????????????205

Glu?Tyr?Pro?His?Val?Pro?Trp?Trp?Gln?Leu?Pro?Glu?Ile?Tyr?Lys?Met

210?????????????????215?????????????????220

Ser?Lys?Ser?Asn?Leu

225

<210>105

<211>1536

<212>DNA

＜213〉the unusual coccus of radiation hardness (Deinococcus radiodurans) R1

<220>

<221>CDS

<222>(1)..(1536)

<223>

atg?ccg?gat?tac?gac?ctg?atc?gtc?atg?ggc?gcg?ggc?cac?aac?gcg?ctg????48

Met?Pro?Asp?Tyr?Asp?Leu?Ile?Val?Met?Gly?Ala?Gly?His?Asn?Ala?Leu

1???????????????5???????????????????10??????????????????15

gtg?act?gct?gcc?tac?gcc?gcc?cgg?gcg?ggc?ctg?aaa?gtc?ggc?gtg?ttc????96

Val?Thr?Ala?Ala?Tyr?Ala?Ala?Arg?Ala?Gly?Leu?Lys?Val?Gly?Val?Phe

20??????????????????25??????????????????30

gag?cgg?cgg?cac?ctc?gtc?ggc?ggg?gcg?gtc?agc?acc?gag?gag?gtc?gtg????144

Glu?Arg?Arg?His?Leu?Val?Gly?Gly?Ala?Val?Ser?Thr?Glu?Glu?Val?Val

35??????????????????40??????????????????45

ccc?ggt?tac?cgc?ttc?gac?tac?ggc?ggc?agc?gcc?cac?atc?ctg?att?cgg????192

Pro?Gly?Tyr?Arg?Phe?Asp?Tyr?Gly?Gly?Ser?Ala?His?Ile?Leu?Ile?Arg

50??????????????????55??????????????????60

atg?acg?ccc?atc?gtg?cgc?gaa?ctc?gaa?ctc?acg?cgg?cac?ggg?ctg?cat????240

Met?Thr?Pro?Ile?Val?Arg?Glu?Leu?Glu?Leu?Thr?Arg?His?Gly?Leu?His

65??????????????????70??????????????????75??????????????????80

tac?ctc?gaa?gtg?gac?cct?atg?ttt?cac?gct?tcc?gac?ggt?gaa?acg?ccc????288

Tyr?Leu?Glu?Val?Asp?Pro?Met?Phe?His?Ala?Ser?Asp?Gly?Glu?Thr?Pro

85??????????????????90??????????????????95

tgg?ttc?att?cac?cgc?gac?gcc?ggg?cgg?acc?atc?cgc?gaa?ctg?gac?gaa????336

Trp?Phe?Ile?His?Arg?Asp?Ala?Gly?Arg?Thr?Ile?Arg?Glu?Leu?Asp?Glu

100?????????????????105?????????????????110

aag?ttt?ccc?ggg?cag?ggc?gac?gcc?tac?ggg?cgc?ttt?ctc?gac?gat?tgg????384

Lys?Phe?Pro?Gly?Gln?Gly?Asp?Ala?Tyr?Gly?Arg?Phe?Leu?Asp?Asp?Trp

115?????????????????120?????????????????125

aca?ccc?ttc?gcg?cgc?gcc?gtg?gcc?gac?ctg?ttc?aac?tcg?gcg?ccg?ggg????432

Thr?Pro?Phe?Ala?Arg?Ala?Val?Ala?Asp?Leu?Phe?Asn?Ser?Ala?Pro?Gly

130?????????????????135?????????????????140

ccg?ctc?gac?ctg?ggc?aaa?atg?gtg?atg?cgc?agc?ggc?cag?ggc?aag?gac????480

Pro?Leu?Asp?Leu?Gly?Lys?Met?Val?Met?Arg?Ser?Gly?Gln?Gly?Lys?Asp

145?????????????????150?????????????????155?????????????????160

tgg?aac?gag?cag?ctc?ccg?cgc?atc?ctg?cgg?ccc?tac?ggc?gac?gtg?gcg????528

Trp?Asn?Glu?Gln?Leu?Pro?Arg?Ile?Leu?Arg?Pro?Tyr?Gly?Asp?Val?Ala

165?????????????????170?????????????????175

cgc?gag?tac?ttc?agc?gag?gag?cgc?gtg?cgg?gct?ccc?ctg?acc?tgg?atg????576

Arg?Glu?Tyr?Phe?Ser?Glu?Glu?Arg?Val?Arg?Ala?Pro?Leu?Thr?Trp?Met

180?????????????????185?????????????????190

gcg?gcc?cag?agc?ggc?ccc?cca?ccc?tcg?gac?ccg?ctg?agc?gcg?ccc?ttt????624

Ala?Ala?Gln?Ser?Gly?Pro?Pro?Pro?Ser?Asp?Pro?Leu?Ser?Ala?Pro?Phe

195?????????????????200?????????????????205

ttg?ctg?tgg?cac?ccg?ctc?tac?cac?gaa?ggc?ggc?gtg?gcg?cgg?ccc?aaa????672

Leu?Leu?Trp?His?Pro?Leu?Tyr?His?Glu?Gly?Gly?Val?Ala?Arg?Pro?Lys

210?????????????????215?????????????????220

ggc?ggc?agc?ggc?ggc?ctg?acc?aaa?gcc?ctg?cgc?cgg?gcc?acc?gag?gcc????720

Gly?Gly?Ser?Gly?Gly?Leu?Thr?Lys?Ala?Leu?Arg?Arg?Ala?Thr?Glu?Ala

225?????????????????230?????????????????235?????????????????240

gaa?ggc?ggc?gag?gtc?ttc?acc?gac?gcg?ccg?gtc?aag?gaa?att?ctg?gtc????768

Glu?Gly?Gly?Glu?Vai?Phe?Thr?Asp?Ala?Pro?Val?Lys?Glu?Ile?Leu?Val

245?????????????????250?????????????????255

aag?gac?ggc?aag?gcg?cag?ggc?atc?cgg?ctg?gaa?agc?ggc?gag?acg?tac????816

Lys?Asp?Gly?Lys?Ala?Gln?Gly?Ile?Arg?Leu?Glu?Ser?Gly?Glu?Thr?Tyr

260?????????????????265?????????????????270

acc?gcc?cgc?gcc?gtc?gtg?tcg?ggc?gtc?cac?atc?ctg?acc?act?gcg?aat????864

Thr?Ala?Arg?Ala?Val?Val?Ser?Gly?Val?His?Ile?Leu?Thr?Thr?Ala?Asn

275?????????????????280?????????????????285

gcc?ctg?ccc?gcc?gaa?tat?gtc?cct?agc?gcc?gcc?agg?aat?gtg?cgc?gtg????912

Ala?Leu?Pro?Ala?Glu?Tyr?Val?Pro?Ser?Ala?Ala?Arg?Asn?Val?Arg?Val

290?????????????????295?????????????????300

ggc?aac?ggc?ttc?ggc?atg?att?ttg?cgc?ctc?gcc?ctc?agt?gaa?aaa?gtc????960

Gly?Asn?Gly?Phe?Gly?Met?Ile?Leu?Arg?Leu?Ala?Leu?Ser?Glu?Lys?Val

305?????????????????310?????????????????315?????????????????320

aaa?tac?cgt?cac?cac?acc?gag?ccc?gac?tca?cgc?atc?ggc?ctg?gga?ttg????1008

Lys?Tyr?Arg?His?His?Thr?Glu?Pro?Asp?Ser?Arg?Ile?Gly?Leu?Gly?Leu

325?????????????????330?????????????????335

ctg?atc?aaa?aac?gag?cgg?caa?atc?atg?cag?ggc?tac?ggc?gaa?tac?ctc????1056

Leu?Ile?Lys?Asn?Glu?Arg?Gln?Ile?Met?Gln?Gly?Tyr?Gly?Glu?Tyr?Leu

340?????????????????345?????????????????350

gcc?ggg?cag?ccc?acc?acc?gac?ccg?ccc?ctc?gtc?gcc?atg?agc?ttc?agc????1104

Ala?Gly?Gln?Pro?Thr?Thr?Asp?Pro?Pro?Leu?Val?Ala?Met?Ser?Phe?Ser

355?????????????????360?????????????????365

gcg?gtg?gac?gac?tcg?ctc?gcc?cca?ccg?aac?ggc?gac?gtg?ttg?tgg?ctg????1152

Ala?Val?Asp?Asp?Ser?Leu?Ala?Pro?Pro?Asn?Gly?Asp?Val?Leu?Trp?Leu

370?????????????????375?????????????????380

tgg?gcg?cag?tac?tac?ccc?ttc?gag?ctc?gcc?acc?ggg?agc?tgg?gaa?acg????1200

Trp?Ala?Gln?Tyr?Tyr?Pro?Phe?Glu?Leu?Ala?Thr?Gly?Ser?Trp?Glu?Thr

385?????????????????390?????????????????395?????????????????400

cgc?acc?gcc?gaa?gcg?cgg?gag?aac?atc?ctg?cgg?gcc?ttt?gag?cac?tac????1248

Arg?Thr?Ala?Glu?Ala?Arg?Glu?Asn?Ile?Leu?Arg?Ala?Phe?Glu?His?Tyr

405?????????????????410?????????????????415

gcg?ccg?ggc?acc?cgc?gac?acg?att?gtg?ggc?gaa?ctc?gtg?cag?acg?ccg????1296

Ala?Pro?Gly?Thr?Arg?Asp?Thr?Ile?Val?Gly?Glu?Leu?Val?Gln?Thr?Pro

420?????????????????425?????????????????430

cag?tgg?ctg?gaa?acc?aac?ctc?ggc?ctg?cac?cgg?ggc?aac?gtg?atg?cac????1344

Gln?Trp?Leu?Glu?Thr?Asn?Leu?Gly?Leu?His?Arg?Gly?Asn?Val?Met?His

435?????????????????440?????????????????445

ctg?gaa?atg?tcc?ttc?gac?cag?atg?ttc?tcc?ttc?cgc?ccc?tgg?ctg?aaa????1392

Leu?Glu?Met?Ser?Phe?Asp?Gln?Met?Phe?Ser?Phe?Arg?Pro?Trp?Leu?Lys

450?????????????????455?????????????????460

gcg?agc?cag?tac?cgc?tgg?ccg?ggc?gtg?cag?ggg?ctg?tac?ctc?acc?ggc????1440

Ala?Ser?Gln?Tyr?Arg?Trp?Pro?Gly?Val?Gln?Gly?Leu?Tyr?Leu?Thr?Gly

465?????????????????470?????????????????475?????????????????480

gcc?agc?acc?cac?ccc?ggc?gga?ggc?atc?atg?ggc?gcc?tcg?gga?cgc?aac????1488

Ala?Ser?Thr?His?Pro?Gly?Gly?Gly?Ile?Met?Gly?Ala?Ser?Gly?Arg?Asn

485?????????????????490?????????????????495

gcg?gcg?cgg?gtc?atc?gtg?aag?gac?ctg?acg?cgg?agg?cgc?tgg?aaa?tga????1536

Ala?Ala?Arg?Val?Ile?Val?Lys?Asp?Leu?Thr?Arg?Arg?Arg?Trp?Lys

500?????????????????505?????????????????510

<210>106

<211>511

<212>PRT

＜213〉the unusual coccus R1 of radiation hardness

<400>106

Met?Pro?Asp?Tyr?Asp?Leu?Ile?Val?Met?Gly?Ala?Gly?His?Asn?Ala?Leu

1???????????????5???????????????????10??????????????????15

Val?Thr?Ala?Ala?Tyr?Ala?Ala?Arg?Ala?Gly?Leu?Lys?Val?Gly?Val?Phe

20??????????????????25??????????????????30

Glu?Arg?Arg?His?Leu?Val?Gly?Gly?Ala?Val?Ser?Thr?Glu?Glu?Val?Val

35??????????????????40??????????????????45

Pro?Gly?Tyr?Arg?Phe?Asp?Tyr?Gly?Gly?Ser?Ala?His?Ile?Leu?Ile?Arg

50??????????????????55??????????????????60

Met?Thr?Pro?Ile?Val?Arg?Glu?Leu?Glu?Leu?Thr?Arg?His?Gly?Leu?His

65??????????????????70??????????????????75??????????????????80

Tyr?Leu?Glu?Val?Asp?Pro?Met?Phe?His?Ala?Ser?Asp?Gly?Glu?Thr?Pro

85??????????????????90??????????????????95

Trp?Phe?Ile?His?Arg?Asp?Ala?Gly?Arg?Thr?Ile?Arg?Glu?Leu?Asp?Glu

100?????????????????105?????????????????110

Lys?Phe?Pro?Gly?Gln?Gly?Asp?Ala?Tyr?Gly?Arg?Phe?Leu?Asp?Asp?Trp

115?????????????????120?????????????????125

Thr?Pro?Phe?Ala?Arg?Ala?Val?Ala?Asp?Leu?Phe?Asn?Ser?Ala?Pro?Gly

130?????????????????135?????????????????140

Pro?Leu?Asp?Leu?Gly?Lys?Met?Val?Met?Arg?Ser?Gly?Gln?Gly?Lys?Asp

145?????????????????150?????????????????155?????????????????160

Trp?Asn?Glu?Gln?Leu?Pro?Arg?Ile?Leu?Arg?Pro?Tyr?Gly?Asp?Val?Ala

165?????????????????170?????????????????175

Arg?Glu?Tyr?Phe?Ser?Glu?Glu?Arg?Val?Arg?Ala?Pro?Leu?Thr?Trp?Met

180?????????????????185?????????????????190

Ala?Ala?Gln?Ser?Gly?Pro?Pro?Pro?Ser?Asp?Pro?Leu?Ser?Ala?Pro?Phe

195?????????????????200?????????????????205

Leu?Leu?Trp?His?Pro?Leu?Tyr?His?Glu?Gly?Gly?Val?Ala?Arg?Pro?Lys

210?????????????????215?????????????????220

Gly?Gly?Ser?Gly?Gly?Leu?Thr?Lys?Ala?Leu?Arg?Arg?Ala?Thr?Glu?Ala

225?????????????????230?????????????????235?????????????????240

Glu?Gly?Gly?Glu?Val?Phe?Thr?Asp?Ala?Pro?Val?Lys?Glu?Ile?Leu?Val

245?????????????????250?????????????????255

Lys?Asp?Gly?Lys?Ala?Gln?Gly?Ile?Arg?Leu?Glu?Ser?Gly?Glu?Thr?Tyr

260?????????????????265?????????????????270

Thr?Ala?Arg?Ala?Val?Val?Ser?Gly?Val?His?Ile?Leu?Thr?Thr?Ala?Asn

275?????????????????280?????????????????285

Ala?Leu?Pro?Ala?Glu?Tyr?Val?Pro?Ser?Ala?Ala?Arg?Asn?Val?Arg?Val

290?????????????????295?????????????????300

Gly?Asn?Gly?Phe?Gly?Met?Ile?Leu?Arg?Leu?Ala?Leu?Ser?Glu?Lys?Val

305?????????????????310?????????????????315?????????????????320

Lys?Tyr?Arg?His?His?Thr?Glu?Pro?Asp?Ser?Arg?Ile?Gly?Leu?Gly?Leu

325?????????????????330?????????????????335

Leu?Ile?Lys?Asn?Glu?Arg?Gln?Ile?Met?Gln?Gly?Tyr?Gly?Glu?Tyr?Leu

340?????????????????345?????????????????350

Ala?Gly?Gln?Pro?Thr?Thr?Asp?Pro?Pro?Leu?Val?Ala?Met?Ser?Phe?Ser

355?????????????????360?????????????????365

Ala?Val?Asp?Asp?Ser?Leu?Ala?Pro?Pro?Asn?Gly?Asp?Val?Leu?Trp?Leu

370?????????????????375?????????????????380

Trp?Ala?Gln?Tyr?Tyr?Pro?Phe?Glu?Leu?Ala?Thr?Gly?Ser?Trp?Glu?Thr

385?????????????????390?????????????????395?????????????????400

Arg?Thr?Ala?Glu?Ala?Arg?Glu?Asn?Ile?Leu?Arg?Ala?Phe?Glu?His?Tyr

405?????????????????410?????????????????415

Ala?Pro?Gly?Thr?Arg?Asp?Thr?Ile?Val?Gly?Glu?Leu?Val?Gln?Thr?Pro

420?????????????????425?????????????????430

Gln?Trp?Leu?Glu?Thr?Asn?Leu?Gly?Leu?His?Arg?Gly?Asn?Val?Met?His

435?????????????????440?????????????????445

Leu?Glu?Met?Ser?Phe?Asp?Gln?Met?Phe?Ser?Phe?Arg?Pro?Trp?Leu?Lys

450?????????????????455?????????????????460

Ala?Ser?Gln?Tyr?Arg?Trp?Pro?Gly?Val?Gln?Gly?Leu?Tyr?Leu?Thr?Gly

465?????????????????470?????????????????475?????????????????480

Ala?Ser?Thr?His?Pro?Gly?Gly?Gly?Ile?Met?Gly?Ala?Ser?Gly?Arg?Asn

485?????????????????490?????????????????495

Ala?Ala?Arg?Val?Ile?Val?Lys?Asp?Leu?Thr?Arg?Arg?Arg?Trp?Lys

500?????????????????505?????????????????510

<210>107

<211>1666

<212>DNA

＜213〉edible tomato (Lycopersicon esculentum)

<220>

<221>CDS

<222>(1)..(1494)

<223>

<400>107

atg?gaa?gct?ctt?ctc?aag?cct?ttt?cca?tct?ctt?tta?ctt?tcc?tct?cct?????48

Met?Glu?Ala?Leu?Leu?Lys?Pro?Phe?Pro?Ser?Leu?Leu?Leu?Ser?Ser?Pro

1???????????????5???????????????????10??????????????????15

aca?ccc?cat?agg?tct?att?ttc?caa?caa?aat?ccc?tct?ttt?cta?agt?ccc?????96

Thr?Pro?His?Arg?Ser?Ile?Phe?Gln?Gln?Asn?Pro?Ser?Phe?Leu?Ser?Pro

20??????????????????25??????????????????30

acc?acc?aaa?aaa?aaa?tca?aga?aaa?tgt?ctt?ctt?aga?aac?aaa?agt?agt????144

Thr?Thr?Lys?Lys?Lys?Ser?Arg?Lys?Cys?Leu?Leu?Arg?Asn?Lys?Ser?Ser

35??????????????????40??????????????????45

aaa?ctt?ttt?tgt?agc?ttt?ctt?gat?tta?gca?ccc?aca?tca?aag?cca?gag????192

Lys?Leu?Phe?Cys?Ser?Phe?Leu?Asp?Leu?Ala?Pro?Thr?Ser?Lys?Pro?Glu

50??????????????????55??????????????????60

tct?tta?gat?gtt?aac?atc?tca?tgg?gtt?gat?cct?aat?tcg?aat?cgg?gct????240

Ser?Leu?Asp?Val?Asn?Ile?Ser?Trp?Val?Asp?Pro?Asn?Ser?Asn?Arg?Ala

65??????????????????70??????????????????75??????????????????80

caa?ttc?gac?gtg?atc?att?atc?gga?gct?ggc?cct?gct?ggg?ctc?agg?cta????288

Gln?Phe?Asp?Val?Ile?Ile?Ile?Gly?Ala?Gly?Pro?Ala?Gly?Leu?Arg?Leu

85??????????????????90??????????????????95

gct?gaa?caa?gtt?tct?aaa?tat?ggt?att?aag?gta?tgt?tgt?gtt?gac?cct????336

Ala?Glu?Gln?Val?Ser?Lys?Tyr?Gly?Ile?Lys?Val?Cys?Cys?Val?Asp?Pro

100?????????????????105?????????????????110

tca?cca?ctc?tcc?atg?tgg?cca?aat?aat?tat?ggt?gtt?tgg?gtt?gat?gag????384

Ser?Pro?Leu?Ser?Met?Trp?Pro?Asn?Asn?Tyr?Gly?Val?Trp?Val?Asp?Glu

115?????????????????120?????????????????125

ttt?gag?aat?tta?gga?ctg?gaa?aat?tgt?tta?gat?cat?aaa?tgg?cct?atg????432

Phe?Glu?Asn?Leu?Gly?Leu?Glu?Asn?Cys?Leu?Asp?His?Lys?Trp?Pro?Met

130?????????????????135?????????????????140

act?tgt?gtg?cat?ata?aat?gat?aac?aaa?act?aag?tat?ttg?gga?aga?cca????480

Thr?Cys?Val?His?Ile?Asn?Asp?Asn?Lys?Thr?Lys?Tyr?Leu?Gly?Arg?Pro

145?????????????????150?????????????????155?????????????????160

tat?ggt?aga?gtt?agt?aga?aag?aag?ctg?aag?ttg?aaa?ttg?ttg?aat?agt????528

Tyr?Gly?Arg?Val?Ser?Arg?Lys?Lys?Leu?Lys?Leu?Lys?Leu?Leu?Asn?Ser

165?????????????????170?????????????????175

tgt?gtt?gag?aac?aga?gtg?aag?ttt?tat?aaa?gct?aag?gtt?tgg?aaa?gtg????576

Cys?Val?Glu?Asn?Arg?Val?Lys?Phe?Tyr?Lys?Ala?Lys?Val?Trp?Lys?Val

180?????????????????185?????????????????190

gaa?cat?gaa?gaa?ttt?gag?tct?tca?att?gtt?tgt?gat?gat?ggt?aag?aag????624

Glu?His?Glu?Glu?Phe?Glu?Ser?Ser?Ile?Val?Cys?Asp?Asp?Gly?Lys?Lys

195?????????????????200?????????????????205

ata?aga?ggt?agt?ttg?gtt?gtg?gat?gca?agt?ggt?ttt?gct?agt?gat?ttt????672

Ile?Arg?Gly?Ser?Leu?Val?Val?Asp?Ala?Ser?Gly?Phe?Ala?Ser?Asp?Phe

210?????????????????215?????????????????220

ata?gag?tat?gac?agg?cca?aga?aac?cat?ggt?tat?caa?att?gct?cat?ggg????720

Ile?Glu?Tyr?Asp?Arg?Pro?Arg?Asn?His?Gly?Tyr?Gln?Ile?Ala?His?Gly

225?????????????????230?????????????????235?????????????????240

gtt?tta?gta?gaa?gtt?gat?aat?cat?cca?ttt?gat?ttg?gat?aaa?atg?gtg????768

Val?Leu?Val?Glu?Val?Asp?Asn?His?Pro?Phe?Asp?Leu?Asp?Lys?Met?Val

245?????????????????250?????????????????255

ctt?atg?gat?tgg?agg?gat?tct?cat?ttg?ggt?aat?gag?cca?tat?tta?agg?????816

Leu?Met?Asp?Trp?Arg?Asp?Ser?His?Leu?Gly?Asn?Glu?Pro?Tyr?Leu?Arg

260?????????????????265?????????????????270

gtg?aat?aat?gct?aaa?gaa?cca?aca?ttc?ttg?tat?gca?atg?cca?ttt?gat?????864

Val?Asn?Asn?Ala?Lys?Glu?Pro?Thr?Phe?Leu?Tyr?Ala?Met?Pro?Phe?Asp

275?????????????????280?????????????????285

aga?gat?ttg?gtt?ttc?ttg?gaa?gag?act?tct?ttg?gtg?agt?cgt?cct?gtt?????912

Arg?Asp?Leu?Val?Phe?Leu?Glu?Glu?Thr?Ser?Leu?Val?Ser?Arg?Pro?Val

290?????????????????295?????????????????300

tta?tcg?tat?atg?gaa?gta?aaa?aga?agg?atg?gtg?gca?aga?tta?agg?cat?????960

Leu?Ser?Tyr?Met?Glu?Val?Lys?Arg?Arg?Met?Val?Ala?Arg?Leu?Arg?His

305?????????????????310?????????????????315?????????????????320

ttg?ggg?atc?aaa?gtg?aaa?agt?gtt?att?gag?gaa?gag?aaa?tgt?gtg?atc????1008

Leu?Gly?Ile?Lys?Val?Lys?Ser?Val?Ile?Glu?Glu?Glu?Lys?Cys?Val?Ile

325?????????????????330?????????????????335

cct?atg?gga?gga?cca?ctt?ccg?cgg?att?cct?caa?aat?gtt?atg?gct?att????1056

Pro?Met?Gly?Gly?Pro?Leu?Pro?Arg?Ile?Pro?Gln?Asn?Val?Met?Ala?Ile

340?????????????????345?????????????????350

ggt?ggg?aat?tca?ggg?ata?gtt?cat?cca?tca?aca?ggg?tac?atg?gtg?gct????1104

Gly?Gly?Asn?Ser?Gly?Ile?Val?His?Pro?Ser?Thr?Gly?Tyr?Met?Val?Ala

355?????????????????360?????????????????365

agg?agc?atg?gct?tta?gca?cca?gta?cta?gct?gaa?gcc?atc?gtc?gag?ggg????1152

Arg?Ser?Met?Ala?Leu?Ala?Pro?Val?Leu?Ala?Glu?Ala?Ile?Val?Glu?Gly

370?????????????????375?????????????????380

ctt?ggc?tca?aca?aga?atg?ata?aga?ggg?tct?caa?ctt?tac?cat?aga?gtt????1200

Leu?Gly?Ser?Thr?Arg?Met?Ile?Arg?Gly?Ser?Gln?Leu?Tyr?His?Arg?Val

385?????????????????390?????????????????395?????????????????400

tgg?aat?ggt?ttg?tgg?cct?ttg?gat?aga?aga?tgt?gtt?aga?gaa?tgt?tat????1248

Trp?Asn?Gly?Leu?Trp?Pro?Leu?Asp?Arg?Arg?Cys?Val?Arg?Glu?Cys?Tyr

405?????????????????410?????????????????415

tca?ttt?ggg?atg?gag?aca?ttg?ttg?aag?ctt?gat?ttg?aaa?ggg?act?agg????1296

Ser?Phe?Gly?Met?Glu?Thr?Leu?Leu?Lys?Leu?Asp?Leu?Lys?Gly?Thr?Arg

420?????????????????425?????????????????430

aga?ttg?ttt?gac?gct?ttc?ttt?gat?ctt?gat?cct?aaa?tac?tgg?caa?ggg????1344

Arg?Leu?Phe?Asp?Ala?Phe?Phe?Asp?Leu?Asp?Pro?Lys?Tyr?Trp?Gln?Gly

435?????????????????440?????????????????445

ttc?ctt?tct?tca?aga?ttg?tct?gtc?aaa?gaa?ctt?ggt?tta?ctc?agc?ttg??????1392

Phe?Leu?Ser?Ser?Arg?Leu?Ser?Val?Lys?Glu?Leu?Gly?Leu?Leu?Ser?Leu

450?????????????????455?????????????????460

tgt?ctt?ttc?gga?cat?ggc?tca?aac?atg?act?agg?ttg?gat?att?gtt?aca??????1440

Cys?Leu?Phe?Gly?His?Gly?Ser?Asn?Met?Thr?Arg?Leu?Asp?Ile?Val?Thr

465?????????????????470?????????????????475?????????????????480

aaa?tgt?cct?ctt?cct?ttg?gtt?aga?ctg?att?ggc?aat?cta?gca?ata?gag??????1488

Lys?Cys?Pro?Leu?Pro?Leu?Val?Arg?Leu?Ile?Gly?Asn?Leu?Ala?Ile?Glu

485?????????????????490?????????????????495

agc?ctt?tgaatgtgaa?aagtttgaat?cattttcttc?attttaattt?ctttgattat???????1544

Ser?Leu

tttcatattt?tctcaattgc?aaaagtgaga?taagagctac?atactgtcaa?caaataaact????1604

actattggaa?agttaaaata?tgtgtttgtt?gtatgttatt?ctaatggaat?ggattttgta????1664

aa???????????????????????????????????????????????????????????????????1666

<210>108

<211>498

<212>PRT

＜213〉edible tomato

<400>108

Met?Glu?Ala?Leu?Leu?Lys?Pro?Phe?Pro?Ser?Leu?Leu?Leu?Ser?Ser?Pro

1???????????????5???????????????????10??????????????????15

Thr?Pro?His?Arg?Ser?Ile?Phe?Gln?Gln?Asn?Pro?Ser?Phe?Leu?Ser?Pro

20??????????????????25??????????????????30

Thr?Thr?Lys?Lys?Lys?Ser?Arg?Lys?Cys?Leu?Leu?Arg?Asn?Lys?Ser?Ser

35??????????????????40??????????????????45

Lys?Leu?Phe?Cys?Ser?Phe?Leu?Asp?Leu?Ala?Pro?Thr?Ser?Lys?Pro?Glu

50??????????????????55??????????????????60

Ser?Leu?Asp?Val?Asn?Ile?Ser?Trp?Val?Asp?Pro?Asn?Ser?Asn?Arg?Ala

65??????????????????70??????????????????75??????????????????80

Gln?Phe?Asp?Val?Ile?Ile?Ile?Gly?Ala?Gly?Pro?Ala?Gly?Leu?Arg?Leu

85??????????????????90??????????????????95

Ala?Glu?Gln?Val?Ser?Lys?Tyr?Gly?Ile?Lys?Val?Cys?Cys?Val?Asp?Pro

100?????????????????105?????????????????110

Ser?Pro?Leu?Ser?Met?Trp?Pro?Asn?Asn?Tyr?Gly?Val?Trp?Val?Asp?Glu

115?????????????????120?????????????????125

Phe?Glu?Asn?Leu?Gly?Leu?Glu?Asn?Cys?Leu?Asp?His?Lys?Trp?Pro?Met

130?????????????????135?????????????????140

Thr?Cys?Val?His?Ile?Asn?Asp?Asn?Lys?Thr?Lys?Tyr?Leu?Gly?Arg?Pro

145?????????????????150?????????????????155?????????????????160

Tyr?Gly?Arg?Val?Ser?Arg?Lys?Lys?Leu?Lys?Leu?Lys?Leu?Leu?Asn?Ser

165?????????????????170?????????????????175

Cys?Val?Glu?Asn?Arg?Val?Lys?Phe?Tyr?Lys?Ala?Lys?Val?Trp?Lys?Val

180?????????????????185?????????????????190

Glu?His?Glu?Glu?Phe?Glu?Ser?Ser?Ile?Val?Cys?Asp?Asp?Gly?Lys?Lys

195?????????????????200?????????????????205

Ile?Arg?Gly?Ser?Leu?Val?Val?Asp?Ala?Ser?Gly?Phe?Ala?Ser?Asp?Phe

210?????????????????215?????????????????220

Ile?Glu?Tyr?Asp?Arg?Pro?Arg?Asn?His?Gly?Tyr?Gln?Ile?Ala?His?Gly

225?????????????????230?????????????????235?????????????????240

Val?Leu?Val?Glu?Val?Asp?Asn?His?Pro?Phe?Asp?Leu?Asp?Lys?Met?Val

245?????????????????250?????????????????255

Leu?Met?Asp?Trp?Arg?Asp?Ser?His?Leu?Gly?Asn?Glu?Pro?Tyr?Leu?Arg

260?????????????????265?????????????????270

Val?Asn?Asn?Ala?Lys?Glu?Pro?Thr?Phe?Leu?Tyr?Ala?Met?Pro?Phe?Asp

275?????????????????280?????????????????285

Arg?Asp?Leu?Val?Phe?Leu?Glu?Glu?Thr?Ser?Leu?Val?Ser?Arg?Pro?Val

290?????????????????295?????????????????300

Leu?Ser?Tyr?Met?Glu?Val?Lys?Arg?Arg?Met?Val?Ala?Arg?Leu?Arg?His

305?????????????????310?????????????????315?????????????????320

Leu?Gly?Ile?Lys?Val?Lys?Ser?Val?Ile?Glu?Glu?Glu?Lys?Cys?Val?Ile

325?????????????????330?????????????????335

Pro?Met?Gly?Gly?Pro?Leu?Pro?Arg?Ile?Pro?Gln?Asn?Val?Met?Ala?Ile

340?????????????????345?????????????????350

Gly?Gly?Asn?Ser?Gly?Ile?Val?His?Pro?Ser?Thr?Gly?Tyr?Met?Val?Ala

355?????????????????360?????????????????365

Arg?Ser?Met?Ala?Leu?Ala?Pro?Val?Leu?Ala?Glu?Ala?Ile?Val?Glu?Gly

370?????????????????375?????????????????380

Leu?Gly?Ser?Thr?Arg?Met?Ile?Arg?Gly?Ser?Gln?Leu?Tyr?His?Arg?Val

385?????????????????390?????????????????395?????????????????400

Trp?Asn?Gly?Leu?Trp?Pro?Leu?Asp?Arg?Arg?Cys?Val?Arg?Glu?Cys?Tyr

405?????????????????410?????????????????415

Ser?Phe?Gly?Met?Glu?Thr?Leu?Leu?Lys?Leu?Asp?Leu?Lys?Gly?Thr?Arg

420?????????????????425?????????????????430

Arg?Leu?Phe?Asp?Ala?Phe?Phe?Asp?Leu?Asp?Pro?Lys?Tyr?Trp?Gln?Gly

435?????????????????440?????????????????445

Phe?Leu?Ser?Ser?Arg?Leu?Ser?Val?Lys?Glu?Leu?Gly?Leu?Leu?Ser?Leu

450?????????????????455?????????????????460

Cys?Leu?Phe?Gly?His?Gly?Ser?Asn?Met?Thr?Arg?Leu?Asp?Ile?Val?Thr

465?????????????????470?????????????????475?????????????????480

Lys?Cys?Pro?Leu?Pro?Leu?Val?Arg?Leu?Ile?Gly?Asn?Leu?Ala?Ile?Glu

485?????????????????490?????????????????495

Ser?Leu

<210>109

<211>1125

<212>DNA

＜213〉edible tomato

<220>

<221>CDS

<222>(20)..(946)

<223>

<400>109

ttggtcatct?ccacaatca?atg?gct?gcc?gcc?gcc?aga?atc?tcc?gcc?tcc?tct????52

Met?Ala?Ala?Ala?Ala?Arg?Ile?Ser?Ala?Ser?Ser

1???????????????5???????????????????10

acc?tca?cga?act?ttt?tat?ttc?cgt?cat?tca?ccg?ttt?ctt?ggc?cca?aaa????100

Thr?Ser?Arg?Thr?Phe?Tyr?Phe?Arg?His?Ser?Pro?Phe?Leu?Gly?Pro?Lys

15??????????????????20??????????????????25

cct?act?tcg?aca?acc?tca?cat?gtt?tct?cca?atc?tct?cct?ttt?tct?ctt????148

Pro?Thr?Ser?Thr?Thr?Ser?His?Val?Ser?Pro?Ile?Ser?Pro?Phe?Ser?Leu

30??????????????????35??????????????????40

aat?cta?ggc?cca?att?ttg?agg?tct?aga?aga?aaa?ccc?agt?ttc?act?gtt????196

Asn?Leu?Gly?Pro?Ile?Leu?Arg?Ser?Arg?Arg?Lys?Pro?Ser?Phe?Thr?Val

45??????????????????50??????????????????55

tgc?ttt?gtt?ctc?gag?gat?gag?aag?ctg?aaa?cct?caa?ttt?gac?gat?gag????244

Cys?Phe?Val?Leu?Glu?Asp?Glu?Lys?Leu?Lys?Pro?Gln?Phe?Asp?Asp?Glu

60??????????????????65??????????????????70??????????????????75

gct?gag?gat?ttt?gaa?aag?aag?att?gag?gaa?cag?atc?tta?gct?act?cgc????292

Ala?Glu?Asp?Phe?Glu?Lys?Lys?Ile?Glu?Glu?Gln?Ile?Leu?Ala?Thr?Arg

80??????????????????85??????????????????90

ttg?gcg?gag?aaa?ctg?gct?agg?aag?aaa?tcg?gag?agg?ttt?act?tat?ctt????340

Leu?Ala?Glu?Lys?Leu?Ala?Arg?Lys?Lys?Ser?Glu?Arg?Phe?Thr?Tyr?Leu

95??????????????????100?????????????????105

gtg?gct?gct?ata?atg?tct?agt?ttt?ggg?att?act?tct?atg?gct?gtt?atg????388

Val?Ala?Ala?Ile?Met?Ser?Ser?Phe?Gly?Ile?Thr?Ser?Met?Ala?Val?Met

110?????????????????115?????????????????120

gct?gtt?tat?tac?aga?ttt?tcg?tgg?caa?atg?gag?gga?gga?gaa?gtt?cct????436

Ala?Val?Tyr?Tyr?Arg?Phe?Ser?Trp?Gln?Met?Glu?Gly?Gly?Glu?Val?Pro

125?????????????????130?????????????????135

gta?acc?gaa?atg?ttg?ggt?aca?ttt?gct?ctc?tct?gtt?ggt?gct?gct?gta????484

Val?Thr?Glu?Met?Leu?Gly?Thr?Phe?Ala?Leu?Ser?Val?Gly?Ala?Ala?Val

140?????????????????145?????????????????150?????????????????155

gga?atg?gag?ttt?tgg?gcg?aga?tgg?gca?cac?aaa?gca?ctg?tgg?cat?gct????532

Gly?Met?Glu?Phe?Trp?Ala?Arg?Trp?Ala?His?Lys?Ala?Leu?Trp?His?Ala

160?????????????????165?????????????????170

tca?cta?tgg?cac?atg?cat?gag?tca?cac?cac?aaa?cca?aga?gaa?gga?cct????580

Ser?Leu?Trp?His?Met?His?Glu?Ser?His?His?Lys?Pro?Arg?Glu?Gly?Pro

175?????????????????180?????????????????185

ttt?gag?ctg?aac?gac?gtt?ttc?gcc?ata?aca?aac?gct?gtt?cca?gca?ata????628

Phe?Glu?Leu?Asn?Asp?Val?Phe?Ala?Ile?Thr?Asn?Ala?Val?Pro?Ala?Ile

190?????????????????195?????????????????200

gcc?ctc?ctc?aac?tat?ggt?ttc?ttc?cat?aaa?ggc?ctc?att?gcc?gga?cta????676

Ala?Leu?Leu?Asn?Tyr?Gly?Phe?Phe?His?Lys?Gly?Leu?Ile?Ala?Gly?Leu

205?????????????????210?????????????????215

tgc?ttc?ggt?gct?ggg?cta?ggg?atc?aca?gta?ttt?gga?atg?gca?tac?atg????724

Cys?Phe?Gly?Ala?Gly?Leu?Gly?Ile?Thr?Val?Phe?Gly?Met?Ala?Tyr?Met

220?????????????????225?????????????????230?????????????????235

ttt?gtt?cac?gat?ggt?ttg?gtt?cac?aag?aga?ttc?cca?gtt?gga?cct?gta????772

Phe?Val?His?Asp?Gly?Leu?Val?His?Lys?Arg?Phe?Pro?Val?Gly?Pro?Val

240?????????????????245?????????????????250

gcc?aat?gta?cct?tat?ctt?agg?aag?gtg?gct?gct?gct?cat?tcg?ctt?cat????820

Ala?Asn?Val?Pro?Tyr?Leu?Arg?Lys?Val?Ala?Ala?Ala?His?Ser?Leu?His

255?????????????????260?????????????????265

cac?tca?gag?aag?ttc?aat?ggt?gtc?cca?tat?ggc?ttg?ttc?ttc?gga?cct????868

His?Ser?Glu?Lys?Phe?Asn?Gly?Val?Pro?Tyr?Gly?Leu?Phe?Phe?Gly?Pro

270?????????????????275?????????????????280

aag?gaa?ctg?gaa?gaa?gta?gga?ggg?acg?gaa?gag?ttg?gaa?aag?gaa?gtg????916

Lys?Glu?Leu?Glu?Glu?Val?Gly?Gly?Thr?Glu?Glu?Leu?Glu?Lys?Glu?Val

285?????????????????290?????????????????295

ata?cga?agg?acg?aga?ctt?tcg?aaa?gga?tca?tgaacgattg?ttcataaaca??????966

Ile?Arg?Arg?Thr?Arg?Leu?Ser?Lys?Gly?Ser

300?????????????????305

tagaatgtca?ttttacactt?cttatcaatg?aggaagggtg?atttttgatg?tatttgatag??1026

tagagaaaaa?tgtagctctc?ttgatgaaat?gaatttgtat?ttatgtaggc?tcttcttatt??1086

cagtaagatt?ttttcttttt?tttgatctcg?tgccgaatt?????????????????????????1125

<210>110

<211>309

<212>PRT

＜213〉edible tomato

<400>110

Met?Ala?Ala?Ala?Ala?Arg?Ile?Ser?Ala?Ser?Ser?Thr?Ser?Arg?Thr?Phe

1???????????????5???????????????????10??????????????????15

Tyr?Phe?Arg?His?Ser?Pro?Phe?Leu?Gly?Pro?Lys?Pro?Thr?Ser?Thr?Thr

20??????????????????25??????????????????30

Ser?His?Val?Ser?Pro?Ile?Ser?Pro?Phe?Ser?Leu?Asn?Leu?Gly?Pro?Ile

35??????????????????40??????????????????45

Leu?Arg?Ser?Arg?Arg?Lys?Pro?Ser?Phe?Thr?Val?Cys?Phe?Val?Leu?Glu

50??????????????????55??????????????????60

Asp?Glu?Lys?Leu?Lys?Pro?Gln?Phe?Asp?Asp?Glu?Ala?Glu?Asp?Phe?Glu

65??????????????????70??????????????????75??????????????????80

Lys?Lys?Ile?Glu?Glu?Gln?Ile?Leu?Ala?Thr?Arg?Leu?Ala?Glu?Lys?Leu

85??????????????????90??????????????????95

Ala?Arg?Lys?Lys?Ser?Glu?Arg?Phe?Thr?Tyr?Leu?Val?Ala?Ala?Ile?Met

100?????????????????105?????????????????110

Ser?Ser?Phe?Gly?Ile?Thr?Ser?Met?Ala?Val?Met?Ala?Val?Tyr?Tyr?Arg

115?????????????????120?????????????????125

Phe?Ser?Trp?Gln?Met?Glu?Gly?Gly?Glu?Val?Pro?Val?Thr?Glu?Met?Leu

130?????????????????135?????????????????140

Gly?Thr?Phe?Ala?Leu?Ser?Val?Gly?Ala?Ala?Val?Gly?Met?Glu?Phe?Trp

145?????????????????150?????????????????155?????????????????160

Ala?Arg?Trp?Ala?His?Lys?Ala?Leu?Trp?His?Ala?Ser?Leu?Trp?His?Met

165?????????????????170?????????????????175

His?Glu?Ser?His?His?Lys?Pro?Arg?Glu?Gly?Pro?Phe?Glu?Leu?Asn?Asp

180?????????????????185?????????????????190

Val?Phe?Ala?Ile?Thr?Asn?Ala?Val?Pro?Ala?Ile?Ala?Leu?Leu?Asn?Tyr

195?????????????????200?????????????????205

Gly?Phe?Phe?His?Lys?Gly?Leu?Ile?Ala?Gly?Leu?Cys?Phe?Gly?Ala?Gly

210?????????????????215?????????????????220

Leu?Gly?Ile?Thr?Val?Phe?Gly?Met?Ala?Tyr?Met?Phe?Val?His?Asp?Gly

225?????????????????230?????????????????235?????????????????240

Leu?Val?His?Lys?Arg?Phe?Pro?Val?Gly?Pro?Val?Ala?Asn?Val?Pro?Tyr

245?????????????????250?????????????????255

Leu?Arg?Lys?Val?Ala?Ala?Ala?His?Ser?Leu?His?His?Ser?Glu?Lys?Phe

260?????????????????265?????????????????270

Asn?Gly?Val?Pro?Tyr?Gly?Leu?Phe?Phe?Gly?Pro?Lys?Glu?Leu?Glu?Glu

275?????????????????280?????????????????285

Val?Gly?Gly?Thr?Glu?Glu?Leu?Glu?Lys?Glu?Val?Ile?Arg?Arg?Thr?Arg

290?????????????????295?????????????????300

Leu?Ser?Lys?Gly?Ser

305

<210>111

<211>1779

<212>DNA

＜213〉mouseearcress

<220>

<221>CDS

<222>(1)..(1779)

<223>

<400>111

atg?gat?ctc?cgt?cgg?agg?cct?cct?aaa?cca?ccg?gtt?acc?aac?aac?aac?????48

Met?Asp?Leu?Arg?Arg?Arg?Pro?Pro?Lys?Pro?Pro?Val?Thr?Asn?Asn?Asn

1???????????????5???????????????????10??????????????????15

aac?tcc?aac?gga?tct?ttc?cgt?tct?tat?cag?cct?cgc?act?tcc?gat?gac?????96

Asn?Ser?Asn?Gly?Ser?Phe?Arg?Ser?Tyr?Gln?Pro?Arg?Thr?Ser?Asp?Asp

20??????????????????25??????????????????30

gat?cat?cgt?cgc?cgg?gct?aca?aca?att?gct?cct?cca?ccg?aaa?gca?tcc????144

Asp?His?Arg?Arg?Arg?Ala?Thr?Thr?Ile?Ala?Pro?Pro?Pro?Lys?Ala?Ser

35??????????????????40??????????????????45

gac?gcg?ctt?cct?ctt?ccg?tta?tat?ctc?aca?aac?gcc?gtt?ttc?ttc?acg????192

Asp?Ala?Leu?Pro?Leu?Pro?Leu?Tyr?Leu?Thr?Asn?Ala?Val?Phe?Phe?Thr

50??????????????????55??????????????????60

ctc?ttc?ttc?tcc?gtc?gcg?tat?tac?ctc?ctc?cac?cgg?tgg?cgt?gac?aag????240

Leu?Phe?Phe?Ser?Val?Ala?Tyr?Tyr?Leu?Leu?His?Arg?Trp?Arg?Asp?Lys

65??????????????????70??????????????????75??????????????????80

atc?cgt?tac?aat?acg?cct?ctt?cac?gtc?gtc?act?atc?aca?gaa?ctc?ggc????288

Ile?Arg?Tyr?Asn?Thr?Pro?Leu?His?Val?Val?Thr?Ile?Thr?Glu?Leu?Gly

85??????????????????90??????????????????95

gcc?att?att?gct?ctc?atc?gct?tcg?ttt?atc?tat?ctc?cta?ggg?ttt?ttt????336

Ala?Ile?Ile?Ala?Leu?Ile?Ala?Ser?Phe?Ile?Tyr?Leu?Leu?Gly?Phe?Phe

100?????????????????105?????????????????110

ggt?att?gac?ttt?gtt?cag?tca?ttt?atc?tca?cgt?gcc?tct?ggt?gat?gct????384

Gly?Ile?Asp?Phe?Val?Gln?Ser?Phe?Ile?Ser?Arg?Ala?Ser?Gly?Asp?Ala

115?????????????????120?????????????????125

tgg?gat?ctc?gcc?gat?acg?atc?gat?gat?gat?gac?cac?cgc?ctt?gtc?acg????432

Trp?Asp?Leu?Ala?Asp?Thr?Ile?Asp?Asp?Asp?Asp?His?Arg?Leu?Val?Thr

130?????????????????135?????????????????140

tgc?tct?cca?ccg?act?ccg?atc?gtt?tcc?gtt?gct?aaa?tta?cct?aat?ccg????480

Cys?Ser?Pro?Pro?Thr?Pro?Ile?Val?Ser?Val?Ala?Lys?Leu?Pro?Asn?Pro

145?????????????????150?????????????????155?????????????????160

gaa?cct?att?gtt?acc?gaa?tcg?ctt?cct?gag?gaa?gac?gag?gag?att?gtg????528

Glu?Pro?Ile?Val?Thr?Glu?Ser?Leu?Pro?Glu?Glu?Asp?Glu?Glu?Ile?Val

165?????????????????170?????????????????175

aaa?tcg?gtt?atc?gac?gga?gtt?att?cca?tcg?tac?tcg?ctt?gaa?tct?cgt?????576

Lys?Ser?Val?Ile?Asp?Gly?Val?Ile?Pro?Ser?Tyr?Ser?Leu?Glu?Ser?Arg

180?????????????????185?????????????????190

ctc?ggt?gat?tgc?aaa?aga?gcg?gcg?tcg?att?cgt?cgt?gag?gcg?ttg?cag?????624

Leu?Gly?Asp?Cys?Lys?Arg?Ala?Ala?Ser?Ile?Arg?Arg?Glu?Ala?Leu?Gln

195?????????????????200?????????????????205

aga?gtc?acc?ggg?aga?tcg?att?gaa?ggg?tta?ccg?ttg?gat?gga?ttt?gat?????672

Arg?Val?Thr?Gly?Arg?Ser?Ile?Glu?Gly?Leu?Pro?Leu?Asp?Gly?Phe?Asp

210?????????????????215?????????????????220

tat?gaa?tcg?att?ttg?ggg?caa?tgc?tgt?gag?atg?cct?gtt?gga?tac?att?????720

Tyr?Glu?Ser?Ile?Leu?Gly?Gln?Cys?Cys?Glu?Met?Pro?Val?Gly?Tyr?Ile

225?????????????????230?????????????????235?????????????????240

cag?att?cct?gtt?ggg?att?gct?ggt?cca?ttg?ttg?ctt?gat?ggt?tat?gag?????768

Gln?Ile?Pro?Val?Gly?Ile?Ala?Gly?Pro?Leu?Leu?Leu?Asp?Gly?Tyr?Glu

245?????????????????250?????????????????255

tac?tct?gtt?cct?atg?gct?aca?acc?gaa?ggt?tgt?ttg?gtt?gct?agc?act?????816

Tyr?Ser?Val?Pro?Met?Ala?Thr?Thr?Glu?Gly?Cys?Leu?Val?Ala?Ser?Thr

260?????????????????265?????????????????270

aac?aga?ggc?tgc?aag?gct?atg?ttt?atc?tct?ggt?ggc?gcc?acc?agt?acc?????864

Asn?Arg?Gly?Cys?Lys?Ala?Met?Phe?Ile?Ser?Gly?Gly?Ala?Thr?Ser?Thr

275?????????????????280?????????????????285

gtt?ctt?aag?gac?ggt?atg?acc?cga?gca?cct?gtt?gtt?cgg?ttc?gct?tcg?????912

Val?Leu?Lys?Asp?Gly?Met?Thr?Arg?Ala?Pro?Val?Val?Arg?Phe?Ala?Ser

290?????????????????295?????????????????300

gcg?aga?cga?gct?tcg?gag?ctt?aag?ttt?ttc?ttg?gag?aat?cca?gag?aac?????960

Ala?Arg?Arg?Ala?Ser?Glu?Leu?Lys?Phe?Phe?Leu?Glu?Asn?Pro?Glu?Asn

305?????????????????310?????????????????315?????????????????320

ttt?gat?act?ttg?gca?gta?gtc?ttc?aac?agg?tcg?agt?aga?ttt?gca?aga????1008

Phe?Asp?Thr?Leu?Ala?Val?Val?Phe?Asn?Arg?Ser?Ser?Arg?Phe?Ala?Arg

325?????????????????330?????????????????335

ctg?caa?agt?gtt?aaa?tgc?aca?atc?gcg?ggg?aag?aat?gct?tat?gta?agg????1056

Leu?Gln?Ser?Val?Lys?Cys?Thr?Ile?Ala?Gly?Lys?Asn?Ala?Tyr?Val?Arg

340?????????????????345?????????????????350

ttc?tgt?tgt?agt?act?ggt?gat?gct?atg?ggg?atg?aat?atg?gtt?tct?aaa????1104

Phe?Cys?Cys?Ser?Thr?Gly?Asp?Ala?Met?Gly?Met?Asn?Met?Val?Ser?Lys

355?????????????????360?????????????????365

ggt?gtg?cag?aat?gtt?ctt?gag?tat?ctt?acc?gat?gat?ttc?cct?gac?atg????1152

Gly?Val?Gln?Asn?Val?Leu?Glu?Tyr?Leu?Thr?Asp?Asp?Phe?Pro?Asp?Met

370?????????????????375?????????????????380

gat?gtg?att?gga?atc?tct?ggt?aac?ttc?tgt?tcg?gac?aag?aaa?cct?gct????1200

Asp?Val?Ile?Gly?Ile?Ser?Gly?Asn?Phe?Cys?Ser?Asp?Lys?Lys?Pro?Ala

385?????????????????390?????????????????395?????????????????400

gct?gtg?aac?tgg?att?gag?gga?cgt?ggt?aaa?tca?gtt?gtt?tgc?gag?gct????1248

Ala?Val?Asn?Trp?Ile?Glu?Gly?Arg?Gly?Lys?Ser?Val?Val?Cys?Glu?Ala

405?????????????????410?????????????????415

gta?atc?aga?gga?gag?atc?gtg?aac?aag?gtc?ttg?aaa?acg?agc?gtg?gct????1296

Val?Ile?Arg?Gly?Glu?Ile?Val?Asn?Lys?Val?Leu?Lys?Thr?Ser?Val?Ala

420?????????????????425?????????????????430

gct?tta?gtc?gag?ctc?aac?atg?ctc?aag?aac?cta?gct?ggc?tct?gct?gtt????1344

Ala?Leu?Val?Glu?Leu?Asn?Met?Leu?Lys?Asn?Leu?Ala?Gly?Ser?Ala?Val

435?????????????????440?????????????????445

gca?ggc?tct?cta?ggt?gga?ttc?aac?gct?cat?gcc?agt?aac?ata?gtg?tct????1392

Ala?Gly?Ser?Leu?Gly?Gly?Phe?Asn?Ala?His?Ala?Ser?Asn?Ile?Val?Ser

450?????????????????455?????????????????460

gct?gta?ttc?ata?gct?act?ggc?caa?gat?cca?gct?caa?aac?gtg?gag?agt????1440

Ala?Val?Phe?Ile?Ala?Thr?Gly?Gln?Asp?Pro?Ala?Gln?Asn?Val?Glu?Ser

465?????????????????470?????????????????475?????????????????480

tct?caa?tgc?atc?acc?atg?atg?gaa?gct?att?aat?gac?ggc?aaa?gat?atc????1488

Ser?Gln?Cys?Ile?Thr?Met?Met?Glu?Ala?Ile?Asn?Asp?Gly?Lys?Asp?Ile

485?????????????????490?????????????????495

cat?atc?tca?gtc?act?atg?cca?tct?atc?gag?gtg?ggg?aca?gtg?gga?gga????1536

His?Ile?Ser?Val?Thr?Met?Pro?Ser?Ile?Glu?Val?Gly?Thr?Val?Gly?Gly

500?????????????????505?????????????????510

gga?aca?cag?ctt?gca?tct?caa?tca?gcg?tgt?tta?aac?ctg?ctc?gga?gtt????1584

Gly?Thr?Gln?Leu?Ala?Ser?Gln?Ser?Ala?Cys?Leu?Asn?Leu?Leu?Gly?Val

515?????????????????520?????????????????525

aaa?gga?gca?agc?aca?gag?tcg?ccg?gga?atg?aac?gca?agg?agg?cta?gcg????1632

Lys?Gly?Ala?Ser?Thr?Glu?Ser?Pro?Gly?Met?Asn?Ala?Arg?Arg?Leu?Ala

530?????????????????535?????????????????540

acg?atc?gta?gcc?gga?gca?gtt?tta?gct?gga?gag?tta?tct?tta?atg?tca????1680

Thr?Ile?Val?Ala?Gly?Ala?Val?Leu?Ala?Gly?Glu?Leu?Ser?Leu?Met?Ser

545?????????????????550?????????????????555?????????????????560

gca?att?gca?gct?gga?cag?ctt?gtg?aga?agt?cac?atg?aaa?tac?aat?aga????1728

Ala?Ile?Ala?Ala?Gly?Gln?Leu?Val?Arg?Ser?His?Met?Lys?Tyr?Asn?Arg

565?????????????????570?????????????????575

tcc?agc?cga?gac?atc?tct?gga?gca?acg?aca?acg?aca?aca?aca?aca?aca????1776

Ser?Ser?Arg?Asp?Ile?Ser?Gly?Ala?Thr?Thr?Thr?Thr?Thr?Thr?Thr?Thr

580?????????????????585?????????????????590

tga????????????????????????????????????????????????????????????????1779

<210>112

<211>592

<212>PRT

＜213〉mouseearcress

<400>112

Met?Asp?Leu?Arg?Arg?Arg?Pro?Pro?Lys?Pro?Pro?Val?Thr?Asn?Asn?Asn

1???????????????5???????????????????10??????????????????15

Asn?Ser?Asn?Gly?Ser?Phe?Arg?Ser?Tyr?Gln?Pro?Arg?Thr?Ser?Asp?Asp

20??????????????????25??????????????????30

Asp?His?Arg?Arg?Arg?Ala?Thr?Thr?Ile?Ala?Pro?Pro?Pro?Lys?Ala?Ser

35??????????????????40??????????????????45

Asp?Ala?Leu?Pro?Leu?Pro?Leu?Tyr?Leu?Thr?Asn?Ala?Val?Phe?Phe?Thr

50??????????????????55??????????????????60

Leu?Phe?Phe?Ser?Val?Ala?Tyr?Tyr?Leu?Leu?His?Arg?Trp?Arg?Asp?Lys

65??????????????????70??????????????????75??????????????????80

Ile?Arg?Tyr?Asn?Thr?Pro?Leu?His?Val?Val?Thr?Ile?Thr?Glu?Leu?Gly

85??????????????????90??????????????????95

Ala?Ile?Ile?Ala?Leu?Ile?Ala?Ser?Phe?Ile?Tyr?Leu?Leu?Gly?Phe?Phe

100?????????????????105?????????????????110

Gly?Ile?Asp?Phe?Val?Gln?Ser?Phe?Ile?Ser?Arg?Ala?Ser?Gly?Asp?Ala

115?????????????????120?????????????????125

Trp?Asp?Leu?Ala?Asp?Thr?Ile?Asp?Asp?Asp?Asp?His?Arg?Leu?Val?Thr

130?????????????????135?????????????????140

Cys?Ser?Pro?Pro?Thr?Pro?Ile?Val?Ser?Val?Ala?Lys?Leu?Pro?Asn?Pro

145?????????????????150?????????????????155?????????????????160

Glu?Pro?Ile?Val?Thr?Glu?Ser?Leu?Pro?Glu?Glu?Asp?Glu?Glu?Ile?Val

165?????????????????170?????????????????175

Lys?Ser?Val?Ile?Asp?Gly?Val?Ile?Pro?Ser?Tyr?Ser?Leu?Glu?Ser?Arg

180?????????????????185?????????????????190

Leu?Gly?Asp?Cys?Lys?Arg?Ala?Ala?Ser?Ile?Arg?Arg?Glu?Ala?Leu?Gln

195?????????????????200?????????????????205

Arg?Val?Thr?Gly?Arg?Ser?Ile?Glu?Gly?Leu?Pro?Leu?Asp?Gly?Phe?Asp

210?????????????????215?????????????????220

Tyr?Glu?Ser?Ile?Leu?Gly?Gln?Cys?Cys?Glu?Met?Pro?Val?Gly?Tyr?Ile

225?????????????????230?????????????????235?????????????????240

Gln?Ile?Pro?Val?Gly?Ile?Ala?Gly?Pro?Leu?Leu?Leu?Asp?Gly?Tyr?Glu

245?????????????????250?????????????????255

Tyr?Ser?Val?Pro?Met?Ala?Thr?Thr?Glu?Gly?Cys?Leu?Val?Ala?Ser?Thr

260?????????????????265?????????????????270

Asn?Arg?Gly?Cys?Lys?Ala?Met?Phe?Ile?Ser?Gly?Gly?Ala?Thr?Ser?Thr

275?????????????????280?????????????????285

Val?Leu?Lys?Asp?Gly?Met?Thr?Arg?Ala?Pro?Val?Val?Arg?Phe?Ala?Ser

290?????????????????295?????????????????300

Ala?Arg?Arg?Ala?Ser?Glu?Leu?Lys?Phe?Phe?Leu?Glu?Asn?Pro?Glu?Asn

305?????????????????310?????????????????315?????????????????320

Phe?Asp?Thr?Leu?Ala?Val?Val?Phe?Asn?Arg?Ser?Ser?Arg?Phe?Ala?Arg

325?????????????????330?????????????????335

Leu?Gln?Ser?Val?Lys?Cys?Thr?Ile?Ala?Gly?Lys?Asn?Ala?Tyr?Val?Arg

340?????????????????345?????????????????350

Phe?Cys?Cys?Ser?Thr?Gly?Asp?Ala?Met?Gly?Met?Asn?Met?Val?Ser?Lys

355?????????????????360?????????????????365

Gly?Val?Gln?Asn?Val?Leu?Glu?Tyr?Leu?Thr?Asp?Asp?Phe?Pro?Asp?Met

370?????????????????375?????????????????380

Asp?Val?Ile?Gly?Ile?Ser?Gly?Asn?phe?Cys?Ser?Asp?Lys?Lys?Pro?Ala

385?????????????????390?????????????????395?????????????????400

Ala?Val?Asn?Trp?Ile?Glu?Gly?Arg?Gly?Lys?Ser?Val?Val?Cys?Glu?Ala

405?????????????????410?????????????????415

Val?Ile?Arg?Gly?Glu?Ile?Val?Asn?Lys?Val?Leu?Lys?Thr?Ser?Val?Ala

420?????????????????425?????????????????430

Ala?Leu?Val?Glu?Leu?Asn?Met?Leu?Lys?Asn?Leu?Ala?Gly?Ser?Ala?Val

435?????????????????440?????????????????445

Ala?Gly?Ser?Leu?Gly?Gly?Phe?Asn?Ala?His?Ala?Ser?Asn?Ile?Val?Ser

450?????????????????455?????????????????460

Ala?Val?Phe?Ile?Ala?Thr?Gly?Gln?Asp?Pro?Ala?Gln?Asn?Val?Glu?Ser

465?????????????????470?????????????????475?????????????????480

Ser?Gln?Cys?Ile?Thr?Met?Met?Glu?Ala?Ile?Asn?Asp?Gly?Lys?Asp?Ile

485?????????????????490?????????????????495

His?Ile?Ser?Val?Thr?Met?Pro?Ser?Ile?Glu?Val?Gly?Thr?Val?Gly?Gly

500?????????????????505?????????????????510

Gly?Thr?Gln?Leu?Ala?Ser?Gln?Ser?Ala?Cys?Leu?Asn?Leu?Leu?Gly?Val

515?????????????????520?????????????????525

Lys?Gly?Ala?Ser?Thr?Glu?Ser?Pro?Gly?Met?Asn?Ala?Arg?Arg?Leu?Ala

530?????????????????535?????????????????540

Thr?Ile?Val?Ala?Gly?Ala?Val?Leu?Ala?Gly?Glu?Leu?Ser?Leu?Met?Ser

545?????????????????550?????????????????555?????????????????560

Ala?Ile?Ala?Ala?Gly?Gln?Leu?Val?Arg?Ser?His?Met?Lys?Tyr?Asn?Arg

565?????????????????570?????????????????575

Ser?Ser?Arg?Asp?Ile?Ser?Gly?Ala?Thr?Thr?Thr?Thr?Thr?Thr?Thr?Thr

580?????????????????585?????????????????590

<210>113

<211>1401

<212>DNA

＜213〉mouseearcress ISPH

<220>

<221>CDS

<222>(1)..(1401)

<223>

<400>113

atg?gct?gtt?gcg?ctc?caa?ttc?agc?cga?tta?tgc?gtt?cga?ccg?gat?act????48

Met?Ala?Val?Ala?Leu?Gln?Phe?Ser?Arg?Leu?Cys?Val?Arg?Pro?Asp?Thr

1???????????????5???????????????????10??????????????????15

ttc?gtg?cgg?gag?aat?cat?ctc?tct?gga?tcc?gga?tct?ctc?cgc?cgc?cgg????96

Phe?Val?Arg?Glu?Asn?His?Leu?Ser?Gly?Ser?Gly?Ser?Leu?Arg?Arg?Arg

20??????????????????25??????????????????30

aaa?gct?tta?tca?gtc?cgg?tgc?tcg?tct?ggc?gat?gag?aac?gct?cct?tcg????144

Lys?Ala?Leu?Ser?Val?Arg?Cys?Ser?Ser?Gly?Asp?Glu?Asn?Ala?Pro?Ser

35??????????????????40??????????????????45

cca?tcg?gtg?gtg?atg?gac?tcc?gat?ttc?gac?gcc?aag?gtg?ttc?cgt?aag????192

Pro?Ser?Val?Val?Met?Asp?Ser?Asp?Phe?Asp?Ala?Lys?Val?Phe?Arg?Lys

50??????????????????55??????????????????60

aac?ttg?acg?aga?agc?gat?aat?tac?aat?cgt?aaa?ggg?ttc?ggt?cat?aag????240

Asn?Leu?Thr?Arg?Ser?Asp?Asn?Tyr?Asn?Arg?Lys?Gly?Phe?Gly?His?Lys

65??????????????????70??????????????????75??????????????????80

gag?gag?aca?ctc?aag?ctc?atg?aat?cga?gag?tac?acc?agt?gat?ata?ttg????288

Glu?Glu?Thr?Leu?Lys?Leu?Met?Asn?Arg?Glu?Tyr?Thr?Ser?Asp?Ile?Leu

85??????????????????90??????????????????95

gag?aca?ctg?aaa?aca?aat?ggg?tat?act?tat?tct?tgg?gga?gat?gtt?act????336

Glu?Thr?Leu?Lys?Thr?Asn?Gly?Tyr?Thr?Tyr?Ser?Trp?Gly?Asp?Val?Thr

100?????????????????105?????????????????110

gtg?aaa?ctc?gct?aaa?gca?tat?ggt?ttt?tgc?tgg?ggt?gtt?gag?cgt?gct????384

Val?Lys?Leu?Ala?Lys?Ala?Tyr?Gly?Phe?Cys?Trp?Gly?Val?Glu?Arg?Ala

115?????????????????120?????????????????125

gtt?cag?att?gca?tat?gaa?gca?cga?aag?cag?ttt?cca?gag?gag?agg?ctt????432

Val?Gln?Ile?Ala?Tyr?Glu?Ala?Arg?Lys?Gln?Phe?Pro?Glu?Glu?Arg?Leu

130?????????????????135?????????????????140

tgg?att?act?aac?gaa?atc?att?cat?aac?ccg?acc?gtc?aat?aag?agg?ttg????480

Trp?Ile?Thr?Asn?Glu?Ile?Ile?His?Asn?Pro?Thr?Val?Asn?Lys?Arg?Leu

145?????????????????150?????????????????155?????????????????160

gaa?gat?atg?gat?gtt?aaa?att?att?ccg?gtt?gag?gat?tca?aag?aaa?cag????528

Glu?Asp?Met?Asp?Val?Lys?Ile?Ile?Pro?Val?Glu?Asp?Ser?Lys?Lys?Gln

165?????????????????170?????????????????175

ttt?gat?gta?gta?gag?aaa?gat?gat?gtg?gtt?atc?ctt?cct?gcg?ttt?gga????576

Phe?Asp?Val?Val?Glu?Lys?Asp?Asp?Val?Val?Ile?Leu?Pro?Ala?Phe?Gly

180?????????????????185?????????????????190

gct?ggt?gtt?gac?gag?atg?tat?gtt?ctt?aat?gat?aaa?aag?gtg?caa?att????624

Ala?Gly?Val?Asp?Glu?Met?Tyr?Val?Leu?Asn?Asp?Lys?Lys?Val?Gln?Ile

195?????????????????200?????????????????205

gtt?gac?acg?act?tgt?cct?tgg?gtg?aca?aag?gtc?tgg?aac?acg?gtt?gag????672

Val?Asp?Thr?Thr?Cys?Pro?Trp?Val?Thr?Lys?Val?Trp?Asn?Thr?Val?Glu

210?????????????????215?????????????????220

aag?cac?aag?aag?ggg?gaa?tac?aca?tca?gta?atc?cat?ggt?aaa?tat?aat????720

Lys?His?Lys?Lys?Gly?Glu?Tyr?Thr?Ser?Val?Ile?His?Gly?Lys?Tyr?Asn

225?????????????????230?????????????????235?????????????????240

cat?gaa?gag?acg?att?gca?act?gcg?tct?ttt?gca?gga?aag?tac?atc?att????768

His?Glu?Glu?Thr?Ile?Ala?Thr?Ala?Ser?Phe?Ala?Gly?Lys?Tyr?Ile?Ile

245?????????????????250?????????????????255

gta?aag?aac?atg?aaa?gag?gca?aat?tac?gtt?tgt?gat?tac?att?ctc?ggt????816

Val?Lys?Asn?Met?Lys?Glu?Ala?Asn?Tyr?Val?Cys?Asp?Tyr?Ile?Leu?Gly

260?????????????????265?????????????????270

ggc?caa?tac?gat?gga?tct?agc?tcc?aca?aaa?gag?gag?ttc?atg?gag?aaa????864

Gly?Gln?Tyr?Asp?Gly?Ser?Ser?Ser?Thr?Lys?Glu?Glu?Phe?Met?Glu?Lys

275?????????????????280?????????????????285

ttc?aaa?tac?gca?att?tcg?aag?ggt?ttc?gat?ccc?gac?aat?gac?ctt?gtc????912

Phe?Lys?Tyr?Ala?Ile?Ser?Lys?Gly?Phe?Asp?Pro?Asp?Asn?Asp?Leu?Val

290?????????????????295?????????????????300

aaa?gtt?ggt?att?gca?aac?caa?aca?acg?atg?cta?aag?gga?gaa?aca?gag????960

Lys?Val?Gly?Ile?Ala?Asn?Gln?Thr?Thr?Met?Leu?Lys?Gly?Glu?Thr?Glu

305?????????????????310?????????????????315?????????????????320

gag?ata?gga?aga?tta?ctc?gag?aca?aca?atg?atg?cgc?aag?tat?gga?gtg????1008

Glu?Ile?Gly?Arg?Leu?Leu?Glu?Thr?Thr?Met?Met?Arg?Lys?Tyr?Gly?Val

325?????????????????330?????????????????335

gaa?aat?gta?agc?gga?cat?ttc?atc?agc?ttc?aac?aca?ata?tgc?gac?gct????1056

Glu?Asn?Val?Ser?Gly?His?Phe?Ile?Ser?Phe?Asn?Thr?Ile?Cys?Asp?Ala

340?????????????????345?????????????????350

act?caa?gag?cga?caa?gac?gca?atc?tat?gag?cta?gtg?gaa?gag?aag?att????1104

Thr?Gln?Glu?Arg?Gln?Asp?Ala?Ile?Tyr?Glu?Leu?Val?Glu?Glu?Lys?Ile

355?????????????????360?????????????????365

gac?ctc?atg?cta?gtg?gtt?ggc?gga?tgg?aat?tca?agt?aac?acc?tct?cac????1152

Asp?Leu?Met?Leu?Val?Val?Gly?Gly?Trp?Asn?Ser?Ser?Asn?Thr?Ser?His

370?????????????????375?????????????????380

ctt?cag?gaa?atc?tca?gag?gca?cgg?gga?atc?cca?tct?tac?tgg?atc?gat????1200

Leu?Gln?Glu?Ile?Ser?Glu?Ala?Arg?Gly?Ile?Pro?Ser?Tyr?Trp?Ile?Asp

385?????????????????390?????????????????395?????????????????400

agt?gag?aaa?cgg?ata?gga?cct?ggg?aat?aaa?ata?gcc?tat?aag?ctc?cac????1248

Ser?Glu?Lys?Arg?Ile?Gly?Pro?Gly?Asn?Lys?Ile?Ala?Tyr?Lys?Leu?His

405?????????????????410?????????????????415

tat?gga?gaa?ctg?gtc?gag?aag?gaa?aac?ttt?ctc?cca?aag?gga?cca?ata????1296

Tyr?Gly?Glu?Leu?Val?Glu?Lys?Glu?Asn?Phe?Leu?Pro?Lys?Gly?Pro?Ile

420?????????????????425?????????????????430

aca?atc?ggt?gtg?aca?tca?ggt?gca?tca?acc?ccg?gat?aag?gtc?gtg?gaa????1344

Thr?Ile?Gly?Val?Thr?Ser?Gly?Ala?Ser?Thr?Pro?Asp?Lys?Val?Val?Glu

435?????????????????440?????????????????445

gat?gct?ttg?gtg?aag?gtg?ttc?gac?att?aaa?cgt?gaa?gag?tta?ttg?cag????1392

Asp?Ala?Leu?Val?Lys?Val?Phe?Asp?Ile?Lys?Arg?Glu?Glu?Leu?Leu?Gln

450?????????????????455?????????????????460

ctg?gct?tga????????????????????????????????????????????????????????1401

Leu?Ala

465

<210>114

<211>466

<212>PRT

＜213〉mouseearcress ISPH

<400>114

Met?Ala?Val?Ala?Leu?Gln?Phe?Ser?Arg?Leu?Cys?Val?Arg?Pro?Asp?Thr

1???????????????5???????????????????10??????????????????15

Phe?Val?Arg?Glu?Asn?His?Leu?Ser?Gly?Ser?Gly?Ser?Leu?Arg?Arg?Arg

20??????????????????25??????????????????30

Lys?Ala?Leu?Ser?Val?Arg?Cys?Ser?Ser?Gly?Asp?Glu?Asn?Ala?Pro?Ser

35??????????????????40??????????????????45

Pro?Ser?Val?Val?Met?Asp?Ser?Asp?Phe?Asp?Ala?Lys?Val?Phe?Arg?Lys

50??????????????????55??????????????????60

Asn?Leu?Thr?Arg?Ser?Asp?Asn?Tyr?Asn?Arg?Lys?Gly?Phe?Gly?His?Lys

65??????????????????70??????????????????75??????????????????80

Glu?Glu?Thr?Leu?Lys?Leu?Met?Asn?Arg?Glu?Tyr?Thr?Ser?Asp?Ile?Leu

85??????????????????90??????????????????95

Glu?Thr?Leu?Lys?Thr?Asn?Gly?Tyr?Thr?Tyr?Ser?Trp?Gly?Asp?Val?Thr

100?????????????????105?????????????????110

Val?Lys?Leu?Ala?Lys?Ala?Tyr?Gly?Phe?Cys?Trp?Gly?Val?Glu?Arg?Ala

115?????????????????120?????????????????125

Val?Gln?Ile?Ala?Tyr?Glu?Ala?Arg?Lys?Gln?Phe?Pro?Glu?Glu?Arg?Leu

130?????????????????135?????????????????140

Trp?Ile?Thr?Asn?Glu?Ile?Ile?His?Asn?Pro?Thr?Val?Asn?Lys?Arg?Leu

145?????????????????150?????????????????155?????????????????160

Glu?Asp?Met?Asp?Val?Lys?Ile?Ile?Pro?Val?Glu?Asp?Ser?Lys?Lys?Gln

165?????????????????170?????????????????175

Phe?Asp?Val?Val?Glu?Lys?Asp?Asp?Val?Val?Ile?Leu?Pro?Ala?Phe?Gly

180?????????????????185?????????????????190

Ala?Gly?Val?Asp?Glu?Met?Tyr?Val?Leu?Asn?Asp?Lys?Lys?Val?Gln?Ile

195?????????????????200?????????????????205

Val?Asp?Thr?Thr?Cys?Pro?Trp?Val?Thr?Lys?Val?Trp?Asn?Thr?Val?Glu

210?????????????????215?????????????????220

Lys?His?Lys?Lys?Gly?Glu?Tyr?Thr?Ser?Val?Ile?His?Gly?Lys?Tyr?Asn

225?????????????????230?????????????????235?????????????????240

His?Glu?Glu?Thr?Ile?Ala?Thr?Ala?Ser?Phe?Ala?Gly?Lys?Tyr?Ile?Ile

245?????????????????250?????????????????255

Val?Lys?Asn?Met?Lys?Glu?Ala?Asn?Tyr?Val?Cys?Asp?Tyr?Ile?Leu?Gly

260?????????????????265?????????????????270

Gly?Gln?Tyr?Asp?Gly?Ser?Ser?Ser?Thr?Lys?Glu?Glu?Phe?Met?Glu?Lys

275?????????????????280?????????????????285

Phe?Lys?Tyr?Ala?Ile?Ser?Lys?Gly?Phe?Asp?Pro?Asp?Asn?Asp?Leu?Val

290?????????????????295?????????????????300

Lys?Val?Gly?Ile?Ala?Asn?Gln?Thr?Thr?Met?Leu?Lys?Gly?Glu?Thr?Glu

305?????????????????310?????????????????315?????????????????320

Glu?Ile?Gly?Arg?Leu?Leu?Glu?Thr?Thr?Met?Met?Arg?Lys?Tyr?Gly?Val

325?????????????????330?????????????????335

Glu?Asn?Val?Ser?Gly?His?Phe?Ile?Ser?Phe?Asn?Thr?Ile?Cys?Asp?Ala

340?????????????????345?????????????????350

Thr?Gln?Glu?Arg?Gln?Asp?Ala?Ile?Tyr?Glu?Leu?Val?Glu?Glu?Lys?Ile

355?????????????????360?????????????????365

Asp?Leu?Met?Leu?Val?Val?Gly?Gly?Trp?Asn?Ser?Ser?Asn?Thr?Ser?His

370?????????????????375?????????????????380

Leu?Gln?Glu?Ile?Ser?Glu?Ala?Arg?Gly?Ile?Pro?Ser?Tyr?Trp?Ile?Asp

385?????????????????390?????????????????395?????????????????400

Ser?Glu?Lys?Arg?Ile?Gly?Pro?Gly?Asn?Lys?Ile?Ala?Tyr?Lys?Leu?His

405?????????????????410?????????????????415

Tyr?Gly?Glu?Leu?Val?Glu?Lys?Glu?Asn?Phe?Leu?Pro?Lys?Gly?Pro?Ile

420?????????????????425?????????????????430

Thr?Ile?Gly?Val?Thr?Ser?Gly?Ala?Ser?Thr?Pro?Asp?Lys?Val?Val?Glu

435?????????????????440?????????????????445

Asp?Ala?Leu?Val?Lys?Val?Phe?Asp?Ile?Lys?Arg?Glu?Glu?Leu?Leu?Gln

450?????????????????455?????????????????460

Leu?Ala

465

<210>115

<211>2160

<212>DNA

＜213〉edible tomato

<220>

<221>CDS

<222>(1)..(2160)

<223>

<400>115

atg?gct?ttg?tgt?gct?tat?gca?ttt?cct?ggg?att?ttg?aac?agg?act?ggt????48

Met?Ala?Leu?Cys?Ala?Tyr?Ala?Phe?Pro?Gly?Ile?Leu?Asn?Arg?Thr?Gly

1???????????????5???????????????????10??????????????????15

gtg?gtt?tca?gat?tct?tct?aag?gca?acc?cct?ttg?ttc?tct?gga?tgg?att????96

Val?Val?Ser?Asp?Ser?Ser?Lys?Ala?Thr?Pro?Leu?Phe?Ser?Gly?Trp?Ile

20??????????????????25??????????????????30

cat?gga?aca?gat?ctg?cag?ttt?ttg?ttc?caa?cac?aag?ctt?act?cat?gag????144

His?Gly?Thr?Asp?Leu?Gln?Phe?Leu?Phe?Gln?His?Lys?Leu?Thr?His?Glu

35??????????????????40??????????????????45

gtc?aag?aaa?agg?tca?cgt?gtg?gtt?cag?gct?tcc?tta?tca?gaa?tct?gga????192

Val?Lys?Lys?Arg?Ser?Arg?Val?Val?Gln?Ala?Ser?Leu?Ser?Glu?Ser?Gly

50??????????????????55??????????????????60

gaa?tac?tac?aca?cag?aga?ccg?cca?acg?cct?att?ttg?gac?act?gtg?aac????240

Glu?Tyr?Tyr?Thr?Gln?Arg?Pro?Pro?Thr?Pro?Ile?Leu?Asp?Thr?Val?Asn

65??????????????????70??????????????????75??????????????????80

tat?ccc?att?cat?atg?aaa?aat?ctg?tct?ctg?aag?gaa?ctt?aaa?caa?cta????288

Tyr?Pro?Ile?His?Met?Lys?Asn?Leu?Ser?Leu?Lys?Glu?Leu?Lys?Gln?Leu

85??????????????????90??????????????????95

gca?gat?gaa?cta?agg?tca?gat?aca?att?ttc?aat?gta?tca?aag?act?ggg????336

Ala?Asp?Glu?Leu?Arg?Ser?Asp?Thr?Ile?Phe?Asn?Val?Ser?Lys?Thr?Gly

100?????????????????105?????????????????110

ggt?cac?ctt?ggc?tca?agt?ctt?ggt?gtt?gtt?gag?ctg?act?gtt?gct?ctt????384

Gly?His?Leu?Gly?Ser?Ser?Leu?Gly?Val?Val?Glu?Leu?Thr?Val?Ala?Leu

115?????????????????120?????????????????125

cat?tat?gtc?ttc?aat?gca?ccg?caa?gat?agg?att?ctc?tgg?gat?gtt?ggt????432

His?Tyr?Val?Phe?Asn?Ala?Pro?Gln?Asp?Arg?Ile?Leu?Trp?Asp?Val?Gly

130?????????????????135?????????????????140

cat?cag?tct?tat?cct?cac?aaa?atc?ttg?act?ggt?aga?agg?gac?aag?atg????480

His?Gln?Ser?Tyr?Pro?His?Lys?Ile?Leu?Thr?Gly?Arg?Arg?Asp?Lys?Met

145?????????????????150?????????????????155?????????????????160

tcg?aca?tta?agg?cag?aca?gat?ggt?ctt?gca?gga?ttt?act?aag?cga?tcg????528

Ser?Thr?Leu?Arg?Gln?Thr?Asp?Gly?Leu?Ala?Gly?Phe?Thr?Lys?Arg?Ser

165?????????????????170?????????????????175

gag?agt?gaa?tat?gat?tgc?ttt?ggc?acc?ggc?cac?agt?tcc?acc?acc?atc????576

Glu?Ser?Glu?Tyr?Asp?Cys?Phe?Gly?Thr?Gly?His?Ser?Ser?Thr?Thr?Ile

180?????????????????185?????????????????190

tca?gca?ggc?cta?ggg?atg?gct?gtt?ggt?aga?gat?cta?aaa?gga?aga?aac????624

Ser?Ala?Gly?Leu?Gly?Met?Ala?Val?Gly?Arg?Asp?Leu?Lys?Gly?Arg?Asn

195?????????????????200?????????????????205

aac?aat?gtt?att?gcc?gta?ata?ggt?gat?ggt?gcc?atg?aca?gca?ggt?caa????672

Asn?Asn?Val?Ile?Ala?Val?Ile?Gly?Asp?Gly?Ala?Met?Thr?Ala?Gly?Gln

210?????????????????215?????????????????220

gct?tat?gaa?gcc?atg?aat?aat?gct?ggt?tac?ctg?gac?tct?gac?atg?att????720

Ala?Tyr?Glu?Ala?Met?Asn?Asn?Ala?Gly?Tyr?Leu?Asp?Ser?Asp?Met?Ile

225?????????????????230?????????????????235?????????????????240

gtt?atc?tta?aac?gac?aat?aga?caa?gtt?tct?tta?cct?act?gct?act?ctg????768

Val?Ile?Leu?Asn?Asp?Asn?Arg?Gln?Val?Ser?Leu?Pro?Thr?Ala?Thr?Leu

245?????????????????250?????????????????255

gat?ggg?cca?gtt?gct?cct?gtt?gga?gct?cta?agt?agt?gct?ttg?agc?agg????816

Asp?Gly?Pro?Val?Ala?Pro?Val?Gly?Ala?Leu?Ser?Ser?Ala?Leu?Ser?Arg

260?????????????????265?????????????????270

tta?cag?tct?aat?agg?cct?ctc?aga?gaa?cta?aga?gaa?gtc?gca?aag?gga????864

Leu?Gln?Ser?Asn?Arg?Pro?Leu?Arg?Glu?Leu?Arg?Glu?Val?Ala?Lys?Gly

275?????????????????280?????????????????285

gtt?act?aag?cag?att?ggt?ggt?cct?atg?cat?gag?ctt?gct?gca?aaa?gtt????912

Val?Thr?Lys?Gln?Ile?Gly?Gly?Pro?Met?His?Glu?Leu?Ala?Ala?Lys?Val

290?????????????????295?????????????????300

gat?gaa?tat?gct?cgt?ggc?atg?att?agt?ggt?tct?gga?tca?aca?ttg?ttt????960

Asp?Glu?Tyr?Ala?Arg?Gly?Met?Ile?Ser?Gly?Ser?Gly?Ser?Thr?Leu?Phe

305?????????????????310?????????????????315?????????????????320

gaa?gaa?ctt?gga?ctt?tac?tat?att?ggt?cct?gtg?gat?ggt?cac?aac?att????1008

Glu?Glu?Leu?Gly?Leu?Tyr?Tyr?Ile?Gly?Pro?Val?Asp?Gly?His?Asn?Ile

325?????????????????330?????????????????335

gat?gat?cta?att?gcg?att?ctc?aaa?gag?gtt?aga?agt?act?aaa?aca?aca????1056

Asp?Asp?Leu?Ile?Ala?Ile?Leu?Lys?Glu?Val?Arg?Ser?Thr?Lys?Thr?Thr

340?????????????????345?????????????????350

ggt?cca?gta?ctg?atc?cat?gtt?gtc?act?gag?aaa?ggc?aga?ggt?tat?cca????1104

Gly?Pro?Val?Leu?Ile?His?Val?Val?Thr?Glu?Lys?Gly?Arg?Gly?Tyr?Pro

355?????????????????360?????????????????365

tat?gct?gag?aga?gct?gca?gat?aag?tat?cat?gga?gtt?gcc?aag?ttt?gat????1152

Tyr?Ala?Glu?Arg?Ala?Ala?Asp?Lys?Tyr?His?Gly?Val?Ala?Lys?Phe?Asp

370?????????????????375?????????????????380

cca?gca?aca?gga?aag?caa?ttc?aaa?gcc?agt?gcc?aag?aca?cag?tcc?tat????1200

Pro?Ala?Thr?Gly?Lys?Gln?Phe?Lys?Ala?Ser?Ala?Lys?Thr?Gln?Ser?Tyr

385?????????????????390?????????????????395?????????????????400

aca?aca?tat?ttt?gcc?gag?gct?tta?att?gca?gaa?gca?gaa?gca?gat?aaa????1248

Thr?Thr?Tyr?Phe?Ala?Glu?Ala?Leu?Ile?Ala?Glu?Ala?Glu?Ala?Asp?Lys

405?????????????????410?????????????????415

gac?att?gtt?gca?atc?cat?gct?gcc?atg?ggg?ggt?ggg?acc?gga?atg?aac????1296

Asp?Ile?Val?Ala?Ile?His?Ala?Ala?Met?Gly?Gly?Gly?Thr?Gly?Met?Asn

420?????????????????425?????????????????430

ctt?ttc?cat?cgt?cgc?ttc?cca?aca?agg?tgt?ttt?gat?gtt?gga?ata?gca????1344

Leu?Phe?His?Arg?Arg?Phe?Pro?Thr?Arg?Cys?Phe?Asp?Val?Gly?Ile?Ala

435?????????????????440?????????????????445

gaa?caa?cat?gca?gta?acc?ttt?gct?gct?gga?ttg?gct?tgt?gaa?ggc?att????1392

Glu?Gln?His?Ala?Val?Thr?Phe?Ala?Ala?Gly?Leu?Ala?Cys?Glu?Gly?Ile

450?????????????????455?????????????????460

aaa?cct?ttc?tgt?gca?atc?tat?tcg?tct?ttc?atg?cag?agg?gct?tat?gac????1440

Lys?Pro?Phe?Cys?Ala?Ile?Tyr?Ser?Ser?Phe?Met?Gln?Arg?Ala?Tyr?Asp

465?????????????????470?????????????????475?????????????????480

cag?gta?gtg?cat?gac?gtt?gat?ttg?caa?aag?ctg?ccc?gtg?agg?ttt?gca????1488

Gln?Val?Val?His?Asp?Val?Asp?Leu?Gln?Lys?Leu?Pro?Val?Arg?Phe?Ala

485?????????????????490?????????????????495

atg?gac?aga?gca?ggt?ctt?gtt?gga?gca?gat?ggt?cca?aca?cat?tgt?ggt????1536

Met?Asp?Arg?Ala?Gly?Leu?Val?Gly?Ala?Asp?Gly?Pro?Thr?His?Cys?Gly

500?????????????????505?????????????????510

gca?ttt?gat?gtt?act?tac?atg?gca?tgt?ctt?cct?aac?atg?gtt?gta?atg????1584

Ala?Phe?Asp?Val?Thr?Tyr?Met?Ala?Cys?Leu?Pro?Asn?Met?Val?Val?Met

515?????????????????520?????????????????525

gct?cct?tct?gat?gaa?gcg?gag?cta?ttt?cac?atg?gta?gca?act?gct?gcc????1632

Ala?Pro?Ser?Asp?Glu?Ala?Glu?Leu?Phe?His?Met?Val?Ala?Thr?Ala?Ala

530?????????????????535?????????????????540

gcc?att?gat?gac?aga?cca?agt?tgt?ttt?aga?tac?cca?aga?gga?aat?ggg????1680

Ala?Ile?Asp?Asp?Arg?Pro?Ser?Cys?Phe?Arg?Tyr?Pro?Arg?Gly?Asn?Gly

545?????????????????550?????????????????555?????????????????560

atc?ggt?gta?gag?ctt?ccg?gct?gga?aac?aaa?gga?att?cct?ctt?gag?gtt????1728

Ile?Gly?Val?Glu?Leu?Pro?Ala?Gly?Asn?Lys?Gly?Ile?Pro?Leu?Glu?Val

565?????????????????570?????????????????575

ggt?aaa?ggt?agg?ata?ttg?att?gag?ggg?gag?aga?gtg?gct?cta?ttg?gga????1776

Gly?Lys?Gly?Arg?Ile?Leu?Ile?Glu?Gly?Glu?Arg?Val?Ala?Leu?Leu?Gly

580?????????????????585?????????????????590

tat?ggc?tca?gca?gtg?cag?aac?tgt?ttg?gat?gct?gct?att?gtg?cta?gaa????1824

Tyr?Gly?Ser?Ala?Val?Gln?Asn?Cys?Leu?Asp?Ala?Ala?Ile?Val?Leu?Glu

595?????????????????600?????????????????605

tcc?cgc?ggc?tta?caa?gta?aca?gtt?gca?gat?gca?cgt?ttc?tgc?aaa?cca????1872

Ser?Arg?Gly?Leu?Gln?Val?Thr?Val?Ala?Asp?Ala?Arg?Phe?Cys?Lys?Pro

610?????????????????615?????????????????620

ctg?gac?cat?gcc?ctc?ata?agg?agc?ctt?gca?aaa?tca?cat?gaa?gtg?cta????1920

Leu?Asp?His?Ala?Leu?Ile?Arg?Ser?Leu?Ala?Lys?Ser?His?Glu?Val?Leu

625?????????????????630?????????????????635?????????????????640

atc?act?gtc?gaa?gaa?gga?tca?att?gga?ggt?ttt?gga?tct?cat?gtt?gtt????1968

Ile?Thr?Val?Glu?Glu?Gly?Ser?Ile?Gly?Gly?Phe?Gly?Ser?His?Val?Val

645?????????????????650?????????????????655

cag?ttc?atg?gcc?tta?gat?ggg?ctt?ctt?gat?ggc?aag?ttg?aag?tgg?aga????2016

Gln?Phe?Met?Ala?Leu?Asp?Gly?Leu?Leu?Asp?Gly?Lys?Leu?Lys?Trp?Arg

660?????????????????665?????????????????670

cca?ata?gtt?ctt?cct?gat?cga?tac?att?gac?cat?gga?tct?cct?gtt?gat????2064

Pro?Ile?Val?Leu?Pro?Asp?Arg?Tyr?Ile?Asp?His?Gly?Ser?Pro?Val?Asp

675?????????????????680?????????????????685

cag?ttg?gcg?gaa?gct?ggc?cta?aca?cca?tct?cac?att?gca?gca?aca?gta????2112

Gln?Leu?Ala?Glu?Ala?Gly?Leu?Thr?Pro?Ser?His?Ile?Ala?Ala?Thr?Val

690?????????????????695?????????????????700

ttt?aac?ata?ctt?gga?caa?acc?aga?gag?gct?cta?gag?gtc?atg?aca?taa????2160

Phe?Asn?Ile?Leu?Gly?Gln?Thr?Arg?Glu?Ala?Leu?Glu?Val?Met?Thr

705?????????????????710?????????????????715

<210>116

<211>719

<212>PRT

＜213〉edible tomato

<400>116

Met?Ala?Leu?Cys?Ala?Tyr?Ala?Phe?Pro?Gly?Ile?Leu?Asn?Arg?Thr?Gly

1???????????????5???????????????????10??????????????????15

Val?Val?Ser?Asp?Ser?Ser?Lys?Ala?Thr?Pro?Leu?Phe?Ser?Gly?Trp?Ile

20??????????????????25??????????????????30

His?Gly?Thr?Asp?Leu?Gln?Phe?Leu?Phe?Gln?His?Lys?Leu?Thr?His?Glu

35??????????????????40??????????????????45

Val?Lys?Lys?Arg?Ser?Arg?Val?Val?Gln?Ala?Ser?Leu?Ser?Glu?Ser?Gly

50??????????????????55??????????????????60

Glu?Tyr?Tyr?Thr?Gln?Arg?Pro?Pro?Thr?Pro?Ile?Leu?Asp?Thr?Val?Asn

65??????????????????70??????????????????75??????????????????80

Tyr?Pro?Ile?His?Met?Lys?Asn?Leu?Ser?Leu?Lys?Glu?Leu?Lys?Gln?Leu

85??????????????????90??????????????????95

Ala?Asp?Glu?Leu?Arg?Ser?Asp?Thr?Ile?Phe?Asn?Val?Ser?Lys?Thr?Gly

100?????????????????105?????????????????110

Gly?His?Leu?Gly?Ser?Ser?Leu?Gly?Val?Val?Glu?Leu?Thr?Val?Ala?Leu

115?????????????????120?????????????????125

His?Tyr?Val?Phe?Asn?Ala?Pro?Gln?Asp?Arg?Ile?Leu?Trp?Asp?Val?Gly

130?????????????????135?????????????????140

His?Gln?Ser?Tyr?Pro?His?Lys?Ile?Leu?Thr?Gly?Arg?Arg?Asp?Lys?Met

145?????????????????150?????????????????155?????????????????160

Ser?Thr?Leu?Arg?Gln?Thr?Asp?Gly?Leu?Ala?Gly?Phe?Thr?Lys?Arg?Ser

165?????????????????170?????????????????175

Glu?Ser?Glu?Tyr?Asp?Cys?Phe?Gly?Thr?Gly?His?Ser?Ser?Thr?Thr?Ile

180?????????????????185?????????????????190

Ser?Ala?Gly?Leu?Gly?Met?Ala?Val?Gly?Arg?Asp?Leu?Lys?Gly?Arg?Asn

195?????????????????200?????????????????205

Asn?Asn?Val?Ile?Ala?Val?Ile?Gly?Asp?Gly?Ala?Met?Thr?Ala?Gly?Gln

210?????????????????215?????????????????220

Ala?Tyr?Glu?Ala?Met?Asn?Asn?Ala?Gly?Tyr?Leu?Asp?Ser?Asp?Met?Ile

225?????????????????230?????????????????235?????????????????240

Val?Ile?Leu?Asn?Asp?Asn?Arg?Gln?Val?Ser?Leu?Pro?Thr?Ala?Thr?Leu

245?????????????????250?????????????????255

Asp?Gly?Pro?Val?Ala?Pro?Val?Gly?Ala?Leu?Ser?Ser?Ala?Leu?Ser?Arg

260?????????????????265?????????????????270

Leu?Gln?Ser?Asn?Arg?Pro?Leu?Arg?Glu?Leu?Arg?Glu?Val?Ala?Lys?Gly

275?????????????????280?????????????????285

Val?Thr?Lys?Gln?Ile?Gly?Gly?Pro?Met?His?Glu?Leu?Ala?Ala?Lys?Val

290?????????????????295?????????????????300

Asp?Glu?Tyr?Ala?Arg?Gly?Met?Ile?Ser?Gly?Ser?Gly?Ser?Thr?Leu?Phe

305?????????????????310?????????????????315?????????????????320

Glu?Glu?Leu?Gly?Leu?Tyr?Tyr?Ile?Gly?Pro?Val?Asp?Gly?His?Asn?Ile

325?????????????????330?????????????????335

Asp?Asp?Leu?Ile?Ala?Ile?Leu?Lys?Glu?Val?Arg?Ser?Thr?Lys?Thr?Thr

340?????????????????345?????????????????350

Gly?Pro?Val?Leu?Ile?His?Val?Val?Thr?Glu?Lys?Gly?Arg?Gly?Tyr?Pro

355?????????????????360?????????????????365

Tyr?Ala?Glu?Arg?Ala?Ala?Asp?Lys?Tyr?His?Gly?Val?Ala?Lys?Phe?Asp

370?????????????????375?????????????????380

Pro?Ala?Thr?Gly?Lys?Gln?Phe?Lys?Ala?Ser?Ala?Lys?Thr?Gln?Ser?Tyr

385?????????????????390?????????????????395?????????????????400

Thr?Thr?Tyr?Phe?Ala?Glu?Ala?Leu?Ile?Ala?Glu?Ala?Glu?Ala?Asp?Lys

405?????????????????410?????????????????415

Asp?Ile?Val?Ala?Ile?His?Ala?Ala?Met?Gly?Gly?Gly?Thr?Gly?Met?Asn

420?????????????????425?????????????????430

Leu?Phe?His?Arg?Arg?Phe?Pro?Thr?Arg?Cys?Phe?Asp?Val?Gly?Ile?Ala

435?????????????????440?????????????????445

Glu?Gln?His?Ala?Val?Thr?Phe?Ala?Ala?Gly?Leu?Ala?Cys?Glu?Gly?Ile

450?????????????????455?????????????????460

Lys?Pro?Phe?Cys?Ala?Ile?Tyr?Ser?Ser?Phe?Met?Gln?Arg?Ala?Tyr?Asp

465?????????????????470?????????????????475?????????????????480

Gln?Val?Val?His?Asp?Val?Asp?Leu?Gln?Lys?Leu?Pro?Val?Arg?Phe?Ala

485?????????????????490?????????????????495

Met?Asp?Arg?Ala?Gly?Leu?Val?Gly?Ala?Asp?Gly?Pro?Thr?His?Cys?Gly

500?????????????????505?????????????????510

Ala?Phe?Asp?Val?Thr?Tyr?Met?Ala?Cys?Leu?Pro?Asn?Met?Val?Val?Met

515?????????????????520?????????????????525

Ala?Pro?Ser?Asp?Glu?Ala?Glu?Leu?Phe?His?Met?Val?Ala?Thr?Ala?Ala

530?????????????????535?????????????????540

Ala?Ile?Asp?Asp?Arg?Pro?Ser?Cys?Phe?Arg?Tyr?Pro?Arg?Gly?Asn?Gly

545?????????????????550?????????????????555?????????????????560

Ile?Gly?Val?Glu?Leu?Pro?Ala?Gly?Asn?Lys?Gly?Ile?Pro?Leu?Glu?Val

565?????????????????570?????????????????575

Gly?Lys?Gly?Arg?Ile?Leu?Ile?Glu?Gly?Glu?Arg?Val?Ala?Leu?Leu?Gly

580?????????????????585?????????????????590

Tyr?Gly?Ser?Ala?Val?Gln?Asn?Cys?Leu?Asp?Ala?Ala?Ile?Val?Leu?Glu

595?????????????????600?????????????????605

Ser?Arg?Gly?Leu?Gln?Val?Thr?Val?Ala?Asp?Ala?Arg?Phe?Cys?Lys?Pro

610?????????????????615?????????????????620

Leu?Asp?His?Ala?Leu?Ile?Arg?Ser?Leu?Ala?Lys?Ser?His?Glu?Val?Leu

625?????????????????630?????????????????635?????????????????640

Ile?Thr?Val?Glu?Glu?Gly?Ser?Ile?Gly?Gly?Phe?Gly?Ser?His?Val?Val

645?????????????????650?????????????????655

Gln?Phe?Met?Ala?Leu?Asp?Gly?Leu?Leu?Asp?Gly?Lys?Leu?Lys?Trp?Arg

660?????????????????665?????????????????670

Pro?Ile?Val?Leu?Pro?Asp?Arg?Tyr?Ile?Asp?His?Gly?Ser?Pro?Val?Asp

675?????????????????680?????????????????685

Gln?Leu?Ala?Glu?Ala?Gly?Leu?Thr?Pro?Ser?His?Ile?Ala?Ala?Thr?Val

690?????????????????695?????????????????700

Phe?Asn?Ile?Leu?Gly?Gln?Thr?Arg?Glu?Ala?Leu?Glu?Val?Met?Thr

705?????????????????710?????????????????715

<210>117

<211>1434

<212>DNA

＜213〉mouseearcress

<220>

<221>CDS

<222>(1)..(1434)

<223>

<400>117

atg?atg?aca?tta?aac?tca?cta?tct?cca?gct?gaa?tcc?aaa?gct?att?tct?????48

Met?Met?Thr?Leu?Asn?Ser?Leu?Ser?Pro?Ala?Glu?Ser?Lys?Ala?Ile?Ser

1???????????????5???????????????????10??????????????????15

ttc?ttg?gat?acc?tcc?agg?ttc?aat?cca?atc?cct?aaa?ctc?tca?ggt?ggg?????96

Phe?Leu?Asp?Thr?Ser?Arg?Phe?Asn?Pro?Ile?Pro?Lys?Leu?Ser?Gly?Gly

20??????????????????25??????????????????30

ttt?agt?ttg?agg?agg?agg?aat?caa?ggg?aga?ggt?ttt?gga?aaa?ggt?gtt????144

Phe?Ser?Leu?Arg?Arg?Arg?Asn?Gln?Gly?Arg?Gly?Phe?Gly?Lys?Gly?Val

35??????????????????40??????????????????45

aag?tgt?tca?gtg?aaa?gtg?cag?cag?caa?caa?caa?cct?cct?cca?gca?tgg????192

Lys?Cys?Ser?Val?Lys?Val?Gln?Gln?Gln?Gln?Gln?Pro?Pro?Pro?Ala?Trp

50??????????????????55??????????????????60

cct?ggg?aga?gct?gtc?cct?gag?gcg?cct?cgt?caa?tct?tgg?gat?gga?cca????240

Pro?Gly?Arg?Ala?Val?Pro?Glu?Ala?Pro?Arg?Gln?Ser?Trp?Asp?Gly?Pro

65??????????????????70??????????????????75??????????????????80

aaa?ccc?atc?tct?atc?gtt?gga?tct?act?ggt?tct?att?ggc?act?cag?aca????288

Lys?Pro?Ile?Ser?Ile?Val?Gly?Ser?Thr?Gly?Ser?Ile?Gly?Thr?Gln?Thr

85??????????????????90??????????????????95

ttg?gat?att?gtg?gct?gag?aat?cct?gac?aaa?ttc?aga?gtt?gtg?gct?cta????336

Leu?Asp?Ile?Val?Ala?Glu?Asn?Pro?Asp?Lys?Phe?Arg?Val?Val?Ala?Leu

100?????????????????105?????????????????110

gct?gct?ggt?tcg?aat?gtt?act?cta?ctt?gct?gat?cag?gta?agg?aga?ttt????384

Ala?Ala?Gly?Ser?Asn?Val?Thr?Leu?Leu?Ala?Asp?Gln?Val?Arg?Arg?Phe

115?????????????????120?????????????????125

aag?cct?gca?ttg?gtt?gct?gtt?aga?aac?gag?tca?ctg?att?aat?gag?ctt????432

Lys?Pro?Ala?Leu?Val?Ala?Val?Arg?Asn?Glu?Ser?Leu?Ile?Asn?Glu?Leu

130?????????????????135?????????????????140

aaa?gag?gct?tta?gct?gat?ttg?gac?tat?aaa?ctc?gag?att?att?cca?gga????480

Lys?Glu?Ala?Leu?Ala?Asp?Leu?Asp?Tyr?Lys?Leu?Glu?Ile?Ile?Pro?Gly

145?????????????????150?????????????????155?????????????????160

gag?caa?gga?gtg?att?gag?gtt?gcc?cga?cat?cct?gaa?gct?gta?acc?gtt????528

Glu?Gln?Gly?Val?Ile?Glu?Val?Ala?Arg?His?Pro?Glu?Ala?Val?Thr?Val

165?????????????????170?????????????????175

gtt?acc?gga?ata?gta?ggt?tgt?gcg?gga?cta?aag?cct?acg?gtt?gct?gca?????576

Val?Thr?Gly?Ile?Val?Gly?Cys?Ala?Gly?Leu?Lys?Pro?Thr?Val?Ala?Ala

180?????????????????185?????????????????190

att?gaa?gca?gga?aag?gac?att?gct?ctt?gca?aac?aaa?gag?aca?tta?atc?????624

Ile?Glu?Ala?Gly?Lys?Asp?Ile?Ala?Leu?Ala?Asn?Lys?Glu?Thr?Leu?Ile

195?????????????????200?????????????????205

gca?ggt?ggt?cct?ttc?gtg?ctt?ccg?ctt?gcc?aac?aaa?cat?aat?gta?aag?????672

Ala?Gly?Gly?Pro?Phe?Val?Leu?Pro?Leu?Ala?Asn?Lys?His?Asn?Val?Lys

210?????????????????215?????????????????220

att?ctt?ccg?gca?gat?tca?gaa?cat?tct?gcc?ata?ttt?cag?tgt?att?caa?????720

Ile?Leu?Pro?Ala?Asp?Ser?Glu?His?Ser?Ala?Ile?Phe?Gln?Cys?Ile?Gln

225?????????????????230?????????????????235?????????????????240

ggt?ttg?cct?gaa?ggc?gct?ctg?cgc?aag?ata?atc?ttg?act?gca?tct?ggt?????768

Gly?Leu?Pro?Glu?Gly?Ala?Leu?Arg?Lys?Ile?Ile?Leu?Thr?Ala?Ser?Gly

245?????????????????250?????????????????255

gga?gct?ttt?agg?gat?tgg?cct?gtc?gaa?aag?cta?aag?gaa?gtt?aaa?gta?????816

Gly?Ala?Phe?Arg?Asp?Trp?Pro?Val?Glu?Lys?Leu?Lys?Glu?Val?Lys?Val

260?????????????????265?????????????????270

gcg?gat?gcg?ttg?aag?cat?cca?aac?tgg?aac?atg?gga?aag?aaa?atc?act?????864

Ala?Asp?Ala?Leu?Lys?His?Pro?Asn?Trp?Asn?Met?Gly?Lys?Lys?Ile?Thr

275?????????????????280?????????????????285

gtg?gac?tct?gct?acg?ctt?ttc?aac?aag?ggt?ctt?gag?gtc?att?gaa?gcg?????912

Val?Asp?Ser?Ala?Thr?Leu?Phe?Asn?Lys?Gly?Leu?Glu?Val?Ile?Glu?Ala

290?????????????????295?????????????????300

cat?tat?ttg?ttt?gga?gct?gag?tat?gac?gat?ata?gag?att?gtc?att?cat?????960

His?Tyr?Leu?Phe?Gly?Ala?Glu?Tyr?Asp?Asp?Ile?Glu?Ile?Val?Ile?His

305?????????????????310?????????????????315?????????????????320

ccg?caa?agt?atc?ata?cat?tcc?atg?att?gaa?aca?cag?gat?tca?tct?gtg????1008

Pro?Gln?Ser?Ile?Ile?His?Ser?Met?Ile?Glu?Thr?Gln?Asp?Ser?Ser?Val

325?????????????????330?????????????????335

ctt?gct?caa?ttg?ggt?tgg?cct?gat?atg?cgt?tta?ccg?att?ctc?tac?acc????1056

Leu?Ala?Gln?Leu?Gly?Trp?Pro?Asp?Met?Arg?Leu?Pro?Ile?Leu?Tyr?Thr

340?????????????????345?????????????????350

atg?tca?tgg?ccc?gat?aga?gtt?cct?tgt?tct?gaa?gta?act?tgg?cca?aga????1104

Met?Ser?Trp?Pro?Asp?Arg?Val?Pro?Cys?Ser?Glu?Val?Thr?Trp?Pro?Arg

355?????????????????360?????????????????365

ctt?gac?ctt?tgc?aaa?ctc?ggt?tca?ttg?act?ttc?aag?aaa?cca?gac?aat????1152

Leu?Asp?Leu?Cys?Lys?Leu?Gly?Ser?Leu?Thr?Phe?Lys?Lys?Pro?Asp?Asn

370?????????????????375?????????????????380

gtg?aaa?tac?cca?tcc?atg?gat?ctt?gct?tat?gct?gct?gga?cga?gct?gga????1200

Val?Lys?Tyr?Pro?Ser?Met?Asp?Leu?Ala?Tyr?Ala?Ala?Gly?Arg?Ala?Gly

385?????????????????390?????????????????395?????????????????400

ggc?aca?atg?act?gga?gtt?ctc?agc?gcc?gcc?aat?gag?aaa?gct?gtt?gaa????1248

Gly?Thr?Met?Thr?Gly?Val?Leu?Ser?Ala?Ala?Asn?Glu?Lys?Ala?Val?Glu

405?????????????????410?????????????????415

atg?ttc?att?gat?gaa?aag?ata?agc?tat?ttg?gat?atc?ttc?aag?gtt?gtg????1296

Met?Phe?Ile?Asp?Glu?Lys?Ile?Ser?Tyr?Leu?Asp?Ile?Phe?Lys?Val?Val

420?????????????????425?????????????????430

gaa?tta?aca?tgc?gat?aaa?cat?cga?aac?gag?ttg?gta?aca?tca?ccg?tct????1344

Glu?Leu?Thr?Cys?Asp?Lys?His?Arg?Asn?Glu?Leu?Val?Thr?Ser?Pro?Ser

435?????????????????440?????????????????445

ctt?gaa?gag?att?gtt?cac?tat?gac?ttg?tgg?gca?cgt?gaa?tat?gcc?gcg????1392

Leu?Glu?Glu?Ile?Val?His?Tyr?Asp?Leu?Trp?Ala?Arg?Glu?Tyr?Ala?Ala

450?????????????????455?????????????????460

aat?gtg?cag?ctt?tct?tct?ggt?gct?agg?cca?gtt?cat?gca?tga????????????1434

Asn?Val?Gln?Leu?Ser?Ser?Gly?Ala?Arg?Pro?Val?His?Ala

465?????????????????470?????????????????475

<210>118

<211>477

<212>PRT

＜213〉mouseearcress

<400>118

Met?Met?Thr?Leu?Asn?Ser?Leu?Ser?Pro?Ala?Glu?Ser?Lys?Ala?Ile?Ser

1???????????????5???????????????????10??????????????????15

Phe?Leu?Asp?Thr?Ser?Arg?Phe?Asn?Pro?Ile?Pro?Lys?Leu?Ser?Gly?Gly

20??????????????????25??????????????????30

Phe?Ser?Leu?Arg?Arg?Arg?Asn?Gln?Gly?Arg?Gly?Phe?Gly?Lys?Gly?Val

35??????????????????40??????????????????45

Lys?Cys?Ser?Val?Lys?Val?Gln?Gln?Gln?Gln?Gln?Pro?Pro?Pro?Ala?Trp

50??????????????????55??????????????????60

Pro?Gly?Arg?Ala?Val?Pro?Glu?Ala?Pro?Arg?Gln?Ser?Trp?Asp?Gly?Pro

65??????????????????70??????????????????75??????????????????80

Lys?Pro?Ile?Ser?Ile?Val?Gly?Ser?Thr?Gly?Ser?Ile?Gly?Thr?Gln?Thr

85??????????????????90??????????????????95

Leu?Asp?Ile?Val?Ala?Glu?Asn?Pro?Asp?Lys?Phe?Arg?Val?Val?Ala?Leu

100?????????????????105?????????????????110

Ala?Ala?Gly?Ser?Asn?Val?Thr?Leu?Leu?Ala?Asp?Gln?Val?Arg?Arg?Phe

115?????????????????120?????????????????125

Lys?Pro?Ala?Leu?Val?Ala?Val?Arg?Asn?Glu?Ser?Leu?Ile?Asn?Glu?Leu

130?????????????????135?????????????????140

Lys?Glu?Ala?Leu?Ala?Asp?Leu?Asp?Tyr?Lys?Leu?Glu?Ile?Ile?Pro?Gly

145?????????????????150?????????????????155?????????????????160

Glu?Gln?Gly?Val?Ile?Glu?Val?Ala?Arg?His?Pro?Glu?Ala?Val?Thr?Val

165?????????????????170?????????????????175

Val?Thr?Gly?Ile?Val?Gly?Cys?Ala?Gly?Leu?Lys?Pro?Thr?Val?Ala?Ala

180?????????????????185?????????????????190

Ile?Glu?Ala?Gly?Lys?Asp?Ile?Ala?Leu?Ala?Asn?Lys?Glu?Thr?Leu?Ile

195?????????????????200?????????????????205

Ala?Gly?Gly?Pro?Phe?Val?Leu?Pro?Leu?Ala?Asn?Lys?His?Asn?Val?Lys

210?????????????????215?????????????????220

Ile?Leu?Pro?Ala?Asp?Ser?Glu?His?Ser?Ala?Ile?Phe?Gln?Cys?Ile?Gln

225?????????????????230?????????????????235?????????????????240

Gly?Leu?Pro?Glu?Gly?Ala?Leu?Arg?Lys?Ile?Ile?Leu?Thr?Ala?Ser?Gly

245?????????????????250?????????????????255

Gly?Ala?Phe?Arg?Asp?Trp?Pro?Val?Glu?Lys?Leu?Lys?Glu?Val?Lys?Val

260?????????????????265?????????????????270

Ala?Asp?Ala?Leu?Lys?His?Pro?Asn?Trp?Asn?Met?Gly?Lys?Lys?Ile?Thr

275?????????????????280?????????????????285

Val?Asp?Ser?Ala?Thr?Leu?Phe?Asn?Lys?Gly?Leu?Glu?Val?Ile?Glu?Ala

290?????????????????295?????????????????300

His?Tyr?Leu?Phe?Gly?Ala?Glu?Tyr?Asp?Asp?Ile?Glu?Ile?Val?Ile?His

305?????????????????310?????????????????315?????????????????320

Pro?Gln?Ser?Ile?Ile?His?Ser?Met?Ile?Glu?Thr?Gln?Asp?Ser?Ser?Val

325?????????????????330?????????????????335

Leu?Ala?Gln?Leu?Gly?Trp?Pro?Asp?Met?Arg?Leu?Pro?Ile?Leu?Tyr?Thr

340?????????????????345?????????????????350

Met?Ser?Trp?Pro?Asp?Arg?Val?Pro?Cys?Ser?Glu?Val?Thr?Trp?Pro?Arg

355?????????????????360?????????????????365

Leu?Asp?Leu?Cys?Lys?Leu?Gly?Ser?Leu?Thr?Phe?Lys?Lys?Pro?Asp?Asn

370?????????????????375?????????????????380

Val?Lys?Tyr?Pro?Ser?Met?Asp?Leu?Ala?Tyr?Ala?Ala?Gly?Arg?Ala?Gly

385?????????????????390?????????????????395?????????????????400

Gly?Thr?Met?Thr?Gly?Val?Leu?Ser?Ala?Ala?Asn?Glu?Lys?Ala?Val?Glu

405?????????????????410?????????????????415

Met?Phe?Ile?Asp?Glu?Lys?Ile?Ser?Tyr?Leu?Asp?Ile?Phe?Lys?Val?Val

420?????????????????425?????????????????430

Glu?Leu?Thr?Cys?Asp?Lys?His?Arg?Asn?Glu?Leu?Val?Thr?Ser?Pro?Ser

435?????????????????440?????????????????445

Leu?Glu?Glu?Ile?Val?His?Tyr?Asp?Leu?Trp?Ala?Arg?Glu?Tyr?Ala?Ala

450?????????????????455?????????????????460

Asn?Val?Gln?Leu?Ser?Ser?Gly?Ala?Arg?Pro?Val?His?Ala

465?????????????????470?????????????????475

<210>119

<211>884

<212>DNA

＜213〉John Burroughs is carried and is received adonis amurensis (Adonis palaestina) clone ApIPI28

<220>

<221>CDS

<222>(180)..(884)

<223>

<400>119

cgtcgatcag?gattaatcct?ttatatagta?tcttctccac?caccactaaa?acattatcag?????60

cttcgtgttc?ttctcccgct?gttcatcttc?agcagcgttg?tcgtactctt?tctatttctt????120

cttccatcac?taacagtcct?cgccgagggt?tgaatcggct?gttcgcctca?acgtcgact?????179

atg?ggt?gaa?gtc?gct?gat?gct?ggt?atg?gat?gcc?gtc?cag?aag?cgg?ctt??????227

Met?Gly?Glu?Val?Ala?Asp?Ala?Gly?Met?Asp?Ala?Val?Gln?Lys?Arg?Leu

1???????????????5???????????????????10??????????????????15

atg?ttc?gac?gat?gaa?tgt?att?ttg?gtg?gat?gag?aat?gac?aag?gtc?gtc????275

Met?Phe?Asp?Asp?Glu?cys?Ile?Leu?Val?Asp?Glu?Asn?Asp?Lys?Val?Val

20??????????????????25??????????????????30

gga?cat?gat?tcc?aaa?tac?aac?tgt?cat?ttg?atg?gaa?aag?ata?gag?gca????323

Gly?His?Asp?Ser?Lys?Tyr?Asn?Cys?His?Leu?Met?Glu?Lys?Ile?Glu?Ala

35??????????????????40??????????????????45

gaa?aac?ttg?ctt?cac?aga?gcc?ttc?agt?gtt?ttc?tta?ttc?aac?tca?aaa????371

Glu?Asn?Leu?Leu?His?Arg?Ala?Phe?Ser?Val?Phe?Leu?Phe?Asn?Ser?Lys

50??????????????????55??????????????????60

tac?gag?ttg?ctt?ctt?cag?caa?cga?tct?gca?acg?aag?gta?aca?ttc?ccg????419

Tyr?Glu?Leu?Leu?Leu?Gln?Gln?Arg?Ser?Ala?Thr?Lys?Val?Thr?Phe?Pro

65??????????????????70??????????????????75??????????????????80

ctc?gta?tgg?aca?aac?acc?tgt?tgc?agc?cat?ccc?ctc?ttc?cgt?gat?tcc????467

Leu?Val?Trp?Thr?Asn?Thr?Cys?Cys?Ser?His?Pro?Leu?Phe?Arg?Asp?Ser

85??????????????????90??????????????????95

gaa?ctc?ata?gaa?gaa?aat?ttt?ctc?ggg?gta?cga?aac?gct?gca?caa?agg????515

Glu?Leu?Ile?Glu?Glu?Asn?Phe?Leu?Gly?Val?Arg?Asn?Ala?Ala?Gln?Arg

100?????????????????105?????????????????110

aag?ctt?tta?gac?gag?cta?ggc?att?cca?gct?gaa?gac?gta?cca?gtt?gat????563

Lys?Leu?Leu?Asp?Glu?Leu?Gly?Ile?Pro?Ala?Glu?Asp?Val?Pro?Val?Asp

115?????????????????120?????????????????125

gaa?ttc?act?cct?ctt?ggt?cgc?att?ctt?tac?aaa?gct?cca?tct?gac?gga????611

Glu?Phe?Thr?Pro?Leu?Gly?Arg?Ile?Leu?Tyr?Lys?Ala?Pro?Ser?Asp?Gly

130?????????????????135?????????????????140

aaa?tgg?gga?gag?cac?gaa?ctg?gac?tat?ctt?ctg?ttt?att?gtc?cga?gat????659

Lys?Trp?Gly?Glu?His?Glu?Leu?Asp?Tyr?Leu?Leu?Phe?Ile?Val?Arg?Asp

145?????????????????150?????????????????155?????????????????160

gtg?aaa?tac?gat?cca?aac?cca?gat?gaa?gtt?gct?gac?gct?aag?tac?gtt????707

Val?Lys?Tyr?Asp?Pro?Asn?Pro?Asp?Glu?Val?Ala?Asp?Ala?Lys?Tyr?Val

165?????????????????170?????????????????175

aat?cgc?gag?gag?ttg?aaa?gag?ata?ctg?aga?aaa?gct?gat?gca?ggt?gaa????755

Asn?Arg?Glu?Glu?Leu?Lys?Glu?Ile?Leu?Arg?Lys?Ala?Asp?Ala?Gly?Glu

180?????????????????185?????????????????190

gag?gga?ata?aag?ttg?tct?cct?tgg?ttt?aga?ttg?gtt?gtg?gat?aac?ttt????803

Glu?Gly?Ile?Lys?Leu?Ser?Pro?Trp?Phe?Arg?Leu?Val?Val?Asp?Asn?Phe

195?????????????????200?????????????????205

ttg?ttc?aag?tgg?tgg?gat?cat?gta?gag?gag?ggg?aag?att?aag?gac?gtc????851

Leu?Phe?Lys?Trp?Trp?Asp?His?Val?Glu?Glu?Gly?Lys?Ile?Lys?Asp?Val

210?????????????????215?????????????????220

gcc?gac?atg?aaa?act?atc?cac?aag?ttg?act?taa????????????????????????884

Ala?Asp?Met?Lys?Thr?Ile?His?Lys?Leu?Thr

225?????????????????230

<210>120

<211>234

<212>PRT

＜213〉John Burroughs is carried and is received adonis amurensis clone ApIPI28

<400>120

Met?Gly?Glu?Val?Ala?Asp?Ala?Gly?Met?Asp?Ala?Val?Gln?Lys?Arg?Leu

1???????????????5???????????????????10??????????????????15

Met?Phe?Asp?Asp?Glu?Cys?Ile?Leu?Val?Asp?Glu?Asn?Asp?Lys?Val?Val

20??????????????????25??????????????????30

Gly?His?Asp?Ser?Lys?Tyr?Asn?Cys?His?Leu?Met?Glu?Lys?Ile?Glu?Ala

35??????????????????40??????????????????45

Glu?Asn?Leu?Leu?His?Arg?Ala?Phe?Ser?Val?Phe?Leu?Phe?Asn?Ser?Lys

50??????????????????55??????????????????60

Tyr?Glu?Leu?Leu?Leu?Gln?Gln?Arg?Ser?Ala?Thr?Lys?Val?Thr?Phe?Pro

65??????????????????70??????????????????75??????????????????80

Leu?Val?Trp?Thr?Asn?Thr?Cys?Cys?Ser?His?Pro?Leu?Phe?Arg?Asp?Ser

85??????????????????90??????????????????95

Glu?Leu?Ile?Glu?Glu?Asn?Phe?Leu?Gly?Val?Arg?Asn?Ala?Ala?Gln?Arg

100?????????????????105?????????????????110

Lys?Leu?Leu?Asp?Glu?Leu?Gly?Ile?Pro?Ala?Glu?Asp?Val?Pro?Val?Asp

115?????????????????120?????????????????125

Glu?Phe?Thr?Pro?Leu?Gly?Arg?Ile?Leu?Tyr?Lys?Ala?Pro?Ser?Asp?Gly

130?????????????????135?????????????????140

Lys?Trp?Gly?Glu?His?Glu?Leu?Asp?Tyr?Leu?Leu?Phe?Ile?Val?Arg?Asp

145?????????????????150?????????????????155?????????????????160

Val?Lys?Tyr?Asp?Pro?Asn?Pro?Asp?Glu?Val?Ala?Asp?Ala?Lys?Tyr?Val

165?????????????????170?????????????????175

Asn?Arg?Glu?Glu?Leu?Lys?Glu?Ile?Leu?Arg?Lys?Ala?Asp?Ala?Gly?Glu

180?????????????????185?????????????????190

Glu?Gly?Ile?Lys?Leu?Ser?Pro?Trp?Phe?Arg?Leu?Val?Val?Asp?Asn?Phe

195?????????????????200?????????????????205

Leu?Phe?Lys?Trp?Trp?Asp?His?Val?Glu?Glu?Gly?Lys?Ile?Lys?Asp?Val

210?????????????????215?????????????????220

Ala?Asp?Met?Lys?Thr?Ile?His?Lys?Leu?Thr

225?????????????????230

<210>121

<211>1402

<212>DNA

＜213〉mouseearcress

<220>

<221>CDS

<222>(52)..(1317)

<223>

<400>121

aagtctttgc?ctctttggtt?tactttcctc?tgttttcgat?ccatttagaa?a?atg?tta????57

Met?Leu

1

ttc?acg?agg?agt?gtt?gct?cgg?att?tct?tct?aag?ttt?ctg?aga?aac?cgt????105

Phe?Thr?Arg?Ser?Val?Ala?Arg?Ile?Ser?Ser?Lys?Phe?Leu?Arg?Asn?Arg

5???????????????????10??????????????????15

agc?ttc?tat?ggc?tcc?tct?caa?tct?ctc?gcc?tct?cat?cgg?ttc?gca?atc????153

Ser?Phe?Tyr?Gly?Ser?Ser?Gln?Ser?Leu?Ala?Ser?His?Arg?Phe?Ala?Ile

20??????????????????25??????????????????30

att?ccc?gat?cag?ggt?cac?tct?tgt?tct?gac?tct?cca?cac?aag?ggt?tac????201

Ile?Pro?Asp?Gln?Gly?His?Ser?Cys?Ser?Asp?Ser?Pro?His?Lys?Gly?Tyr

35??????????????????40??????????????????45??????????????????50

gtt?tgc?aga?aca?act?tat?tca?ttg?aaa?tct?ccg?gtt?ttt?ggt?gga?ttt????249

Val?Cys?Arg?Thr?Thr?Tyr?Ser?Leu?Lys?Ser?Pro?Val?Phe?Gly?Gly?Phe

55??????????????????60??????????????????65

agt?cat?caa?ctc?tat?cac?cag?agt?agc?tcc?ttg?gtt?gag?gag?gag?ctt????297

Ser?His?Gln?Leu?Tyr?His?Gln?Ser?Ser?Ser?Leu?Val?Glu?Glu?Glu?Leu

70??????????????????75??????????????????80

gac?cca?ttt?tcg?ctt?gtt?gcc?gat?gag?ctg?tca?ctt?ctt?agt?aat?aag????345

Asp?Pro?Phe?Ser?Leu?Val?Ala?Asp?Glu?Leu?Ser?Leu?Leu?Ser?Asn?Lys

85??????????????????90??????????????????95

ttg?aga?gag?atg?gta?ctt?gcc?gag?gtt?cca?aag?ctt?gcc?tct?gct?gct????393

Leu?Arg?Glu?Met?Val?Leu?Ala?Glu?Val?Pro?Lys?Leu?Ala?Ser?Ala?Ala

100?????????????????105?????????????????110

gag?tac?ttc?ttc?aaa?agg?ggt?gtg?caa?gga?aaa?cag?ttt?cgt?tca?act????441

Glu?Tyr?Phe?Phe?Lys?Arg?Gly?Val?Gln?Gly?Lys?Gln?Phe?Arg?Ser?Thr

115?????????????????120?????????????????125?????????????????130

att?ttg?ctg?ctg?atg?gcg?aca?gct?ctg?gat?gta?cga?gtt?cca?gaa?gca????489

Ile?Leu?Leu?Leu?Met?Ala?Thr?Ala?Leu?Asp?Val?Arg?Val?Pro?Glu?Ala

135?????????????????140?????????????????145

ttg?att?ggg?gaa?tca?aca?gat?ata?gtc?aca?tca?gaa?tta?cgc?gta?agg?????537

Leu?Ile?Gly?Glu?Ser?Thr?Asp?Ile?Val?Thr?Ser?Glu?Leu?Arg?Val?Arg

150?????????????????155?????????????????160

caa?cgg?ggt?att?gct?gaa?atc?act?gaa?atg?ata?cac?gtc?gca?agt?cta?????585

Gln?Arg?Gly?Ile?Ala?Glu?Ile?Thr?Glu?Met?Ile?His?Val?Ala?Ser?Leu

165?????????????????170?????????????????165

ctg?cac?gat?gat?gtc?ttg?gat?gat?gcc?gat?aca?agg?cgt?ggt?gtt?ggt?????633

Leu?His?Asp?Asp?Val?Leu?Asp?Asp?Ala?Asp?Thr?Arg?Arg?Gly?Val?Gly

180?????????????????185?????????????????190

tcc?tta?aat?gtt?gta?atg?ggt?aac?aag?atg?tcg?gta?tta?gca?gga?gac?????681

Ser?Leu?Asn?Val?Val?Met?Gly?Asn?Lys?Met?Ser?Val?Leu?Ala?Gly?Asp

195?????????????????200?????????????????205?????????????????210

ttc?ttg?ctc?tcc?cgg?gct?tgt?ggg?gct?ctc?gct?gct?tta?aag?aac?aca?????729

Phe?Leu?Leu?Ser?Arg?Ala?Cys?Gly?Ala?Leu?Ala?Ala?Leu?Lys?Asn?Thr

215?????????????????220?????????????????225

gag?gtt?gta?gca?tta?ctt?gca?act?gct?gta?gaa?cat?ctt?gtt?acc?ggt?????777

Glu?Val?Val?Ala?Leu?Leu?Ala?Thr?Ala?Val?Glu?His?Leu?Val?Thr?Gly

230?????????????????235?????????????????240

gaa?acc?atg?gag?ata?act?agt?tca?acc?gag?cag?cgt?tat?agt?atg?gac?????825

Glu?Thr?Met?Glu?Ile?Thr?Ser?Ser?Thr?Glu?Gln?Arg?Tyr?Ser?Met?Asp

245?????????????????250?????????????????255

tac?tac?atg?cag?aag?aca?tat?tat?aag?aca?gca?tcg?cta?atc?tct?aac?????873

Tyr?Tyr?Met?Gln?Lys?Thr?Tyr?Tyr?Lys?Thr?Ala?Ser?Leu?Ile?Ser?Asn

260?????????????????265?????????????????270

agc?tgc?aaa?gct?gtt?gcc?gtt?ctc?act?gga?caa?aca?gca?gaa?gtt?gcc?????921

Ser?Cys?Lys?Ala?Val?Ala?Val?Leu?Thr?Gly?Gln?Thr?Ala?Glu?Val?Ala

275?????????????????280?????????????????285?????????????????290

gtg?tta?gct?ttt?gag?tat?ggg?agg?aat?ctg?ggt?tta?gca?ttc?caa?tta?????969

Val?Leu?Ala?Phe?Glu?Tyr?Gly?Arg?Asn?Leu?Gly?Leu?Ala?Phe?Gln?Leu

295?????????????????300?????????????????305

ata?gac?gac?att?ctt?gat?ttc?acg?ggc?aca?tct?gcc?tct?ctc?gga?aag????1017

Ile?Asp?Asp?Ile?Leu?Asp?Phe?Thr?Gly?Thr?Ser?Ala?Ser?Leu?Gly?Lys

310?????????????????315?????????????????320

gga?tcg?ttg?tca?gat?att?cgc?cat?gga?gtc?ata?aca?gcc?cca?atc?ctc????1065

Gly?Ser?Leu?Ser?Asp?Ile?Arg?His?Gly?Val?Ile?Thr?Ala?Pro?Ile?Leu

325?????????????????330?????????????????335

ttt?gcc?atg?gaa?gag?ttt?cct?caa?cta?cgc?gaa?gtt?gtt?gat?caa?gtt????1113

Phe?Ala?Met?Glu?Glu?Phe?Pro?Gln?Leu?Arg?Glu?Val?Val?Asp?Gln?Val

340?????????????????345?????????????????350

gaa?aaa?gat?cct?agg?aat?gtt?gac?att?gct?tta?gag?tat?ctt?ggg?aag????1161

Glu?Lys?Asp?Pro?Arg?Asn?Val?Asp?Ile?Ala?Leu?Glu?Tyr?Leu?Gly?Lys

355?????????????????360?????????????????365?????????????????370

agc?aag?gga?ata?cag?agg?gca?aga?gaa?tta?gcc?atg?gaa?cat?gcg?aat????1209

Ser?Lys?Gly?Ile?Gln?Arg?Ala?Arg?Glu?Leu?Ala?Met?Glu?His?Ala?Asn

375?????????????????380?????????????????385

cta?gca?gca?gct?gca?atc?ggg?tct?cta?cct?gaa?aca?gac?aat?gaa?gat????1257

Leu?Ala?Ala?Ala?Ala?Ile?Gly?Ser?Leu?Pro?Glu?Thr?Asp?Asn?Glu?Asp

390?????????????????395?????????????????400

gtc?aaa?aga?tcg?agg?cgg?gca?ctt?att?gac?ttg?acc?cat?aga?gtc?atc????1305

Val?Lys?Arg?Ser?Arg?Arg?Ala?Leu?Ile?Asp?Leu?Thr?His?Arg?Val?Ile

405?????????????????410?????????????????415

acc?aga?aac?aag?tgagattaag?taatgtttct?ctctatacac?caaaacattc????????1357

Thr?Arg?Asn?Lys

420

ctcatttcat?ttgtaggatt?ttgttggtcc?aattcgtttc?acgaa??????????????????1402

<210>122

<211>422

<212>PRT

＜213〉mouseearcress

<400>122

Met?Leu?Phe?Thr?Arg?Ser?Val?Ala?Arg?Ile?Ser?Ser?Lys?Phe?Leu?Arg

1???????????????5???????????????????10??????????????????15

Asn?Arg?Ser?Phe?Tyr?Gly?Ser?Ser?Gln?Ser?Leu?Ala?Ser?His?Arg?Phe

20??????????????????25??????????????????30

Ala?Ile?Ile?Pro?Asp?Gln?Gly?His?Ser?Cys?Ser?Asp?Ser?Pro?His?Lys

35??????????????????40??????????????????45

Gly?Tyr?Val?Cys?Arg?Thr?Thr?Tyr?Ser?Leu?Lys?Ser?Pro?Val?Phe?Gly

50??????????????????55??????????????????60

Gly?Phe?Ser?His?Gln?Leu?Tyr?His?Gln?Ser?Ser?Ser?Leu?Val?Glu?Glu

65??????????????????70??????????????????75??????????????????80

Glu?Leu?Asp?Pro?Phe?Ser?Leu?Val?Ala?Asp?Glu?Leu?Ser?Leu?Leu?Ser

85??????????????????90??????????????????95

Asn?Lys?Leu?Arg?Glu?Met?Val?Leu?Ala?Glu?Val?Pro?Lys?Leu?Ala?Ser

100?????????????????105?????????????????110

Ala?Ala?Glu?Tyr?Phe?Phe?Lys?Arg?Gly?Val?Gln?Gly?Lys?Gln?Phe?Arg

115?????????????????120?????????????????125

Ser?Thr?Ile?Leu?Leu?Leu?Met?Ala?Thr?Ala?Leu?Asp?Val?Arg?Val?Pro

130?????????????????135?????????????????140

Glu?Ala?Leu?Ile?Gly?Glu?Ser?Thr?Asp?Ile?Val?Thr?Ser?Glu?Leu?Arg

145?????????????????150?????????????????155?????????????????160

Val?Arg?Gln?Arg?Gly?Ile?Ala?Glu?Ile?Thr?Glu?Met?Ile?His?Val?Ala

165?????????????????170?????????????????175

Ser?Leu?Leu?His?Asp?Asp?Val?Leu?Asp?Asp?Ala?Asp?Thr?Arg?Arg?Gly

180?????????????????185?????????????????190

Val?Gly?Ser?Leu?Asn?Val?Val?Met?Gly?Asn?Lys?Met?Ser?Val?Leu?Ala

195?????????????????200?????????????????205

Gly?Asp?Phe?Leu?Leu?Ser?Arg?Ala?Cys?Gly?Ala?Leu?Ala?Ala?Leu?Lys

210?????????????????215?????????????????220

Asn?Thr?Glu?Val?Val?Ala?Leu?Leu?Ala?Thr?Ala?Val?Glu?His?Leu?Val

225?????????????????230?????????????????235?????????????????24

Thr?Gly?Glu?Thr?Met?Glu?Ile?Thr?Ser?Ser?Thr?Glu?Gln?Arg?Tyr?Ser

245?????????????????250?????????????????255

Met?Asp?Tyr?Tyr?Met?Gln?Lys?Thr?Tyr?Tyr?Lys?Thr?Ala?Ser?Leu?Ile

260?????????????????265?????????????????270

Ser?Asn?Ser?Cys?Lys?Ala?Val?Ala?Val?Leu?Thr?Gly?Gln?Thr?Ala?Glu

275?????????????????280?????????????????285

Val?Ala?Val?Leu?Ala?Phe?Glu?Tyr?Gly?Arg?Asn?Leu?Gly?Leu?Ala?Phe

290?????????????????295?????????????????300

Gln?Leu?Ile?Asp?Asp?Ile?Leu?Asp?Phe?Thr?Gly?Thr?Ser?Ala?Ser?Leu

305?????????????????310?????????????????315?????????????????320

Gly?Lys?Gly?Ser?Leu?Ser?Asp?Ile?Arg?His?Gly?Val?Ile?Thr?Ala?Pro

325?????????????????330?????????????????335

Ile?Leu?Phe?Ala?Met?Glu?Glu?Phe?Pro?Gln?Leu?Arg?Glu?Val?Val?Asp

340?????????????????345?????????????????350

Gln?Val?Glu?Lys?Asp?Pro?Arg?Asn?Val?Asp?Ile?Ala?Leu?Glu?Tyr?Leu

355?????????????????360?????????????????365

Gly?Lys?Ser?Lys?Gly?Ile?Gln?Arg?Ala?Arg?Glu?Leu?Ala?Met?Glu?His

370?????????????????375?????????????????380

Ala?Asn?Leu?Ala?Ala?Ala?Ala?Ile?Gly?Ser?Leu?Pro?Glu?Thr?Asp?Asn

385?????????????????390?????????????????395?????????????????400

Glu?Asp?Val?Lys?Arg?Ser?Arg?Arg?Ala?Leu?Ile?Asp?Leu?Thr?His?Arg

405?????????????????410????????????????415

Val?Ile?Thr?Arg?Asn?Lys

420

<210>123

<211>1155

<212>DNA

＜213〉mouseearcress

<220>

<221>CDS

<222>(1)..(1155)

<223>

<400>123

atg?agt?gtg?agt?tgt?tgt?tgt?agg?aat?ctg?ggc?aag?aca?ata?aaa?aag????48

Met?Ser?Val?Ser?Cys?Cys?Cys?Arg?Asn?Leu?Gly?Lys?Thr?Ile?Lys?Lys

1???????????????5???????????????????10??????????????????15

gca?ata?cct?tca?cat?cat?ttg?cat?ctg?aga?agt?ctt?ggt?ggg?agt?ctc????96

Ala?Ile?Pro?Ser?His?His?Leu?His?Leu?Arg?Ser?Leu?Gly?Gly?Ser?Leu

20??????????????????25??????????????????30

tat?cgt?cgt?cgt?atc?caa?agc?tct?tca?atg?gag?acc?gat?ctc?aag?tca????144

Tyr?Arg?Arg?Arg?Ile?Gln?Ser?Ser?Ser?Met?Glu?Thr?Asp?Leu?Lys?Ser

35??????????????????40??????????????????45

acc?ttt?ctc?aac?gtt?tat?tct?gtt?ctc?aag?tct?gac?ctt?ctt?cat?gac????192

Thr?Phe?Leu?Asn?Val?Tyr?Ser?Val?Leu?Lys?Ser?Asp?Leu?Leu?His?Asp

50??????????????????55??????????????????60

cct?tcc?ttc?gaa?ttc?acc?aat?gaa?tct?cgt?ctc?tgg?gtt?gat?cgg?atg????240

Pro?Ser?Phe?Glu?Phe?Thr?Asn?Glu?Ser?Arg?Leu?Trp?Val?Asp?Arg?Met

65??????????????????70??????????????????75??????????????????80

ctg?gac?tac?aat?gta?cgt?gga?ggg?aaa?ctc?aat?cgg?ggt?ctc?tct?gtt????288

Leu?Asp?Tyr?Asn?Val?Arg?Gly?Gly?Lys?Leu?Asn?Arg?Gly?Leu?Ser?Val

85??????????????????90??????????????????95

gtt?gac?agt?ttc?aaa?ctt?ttg?aag?caa?ggc?aat?gat?ttg?act?gag?caa????336

Val?Asp?Ser?Phe?Lys?Leu?Leu?Lys?Gln?Gly?Asn?Asp?Leu?Thr?Glu?Gln

100?????????????????105?????????????????110

gag?gtt?ttc?ctc?tct?tgt?gct?ctc?ggt?tgg?tgc?att?gaa?tgg?ctc?caa????384

Glu?Val?Phe?Leu?Ser?Cys?Ala?Leu?Gly?Trp?Cys?Ile?Glu?Trp?Leu?Gln

115?????????????????120?????????????????125

gct?tat?ttc?ctt?gtg?ctt?gat?gat?att?atg?gat?aac?tct?gtc?act?cgc????432

Ala?Tyr?Phe?Leu?Val?Leu?Asp?Asp?Ile?Met?Asp?Asn?Ser?Val?Thr?Arg

130?????????????????135?????????????????140

cgt?ggt?caa?cct?tgc?tgg?ttc?aga?gtt?cct?cag?gtt?ggt?atg?gtt?gcc????480

Arg?Gly?Gln?Pro?Cys?Trp?Phe?Arg?Val?Pro?Gln?Val?Gly?Met?Val?Ala

145?????????????????150?????????????????155?????????????????160

atc?aat?gat?ggg?att?cta?ctt?cgc?aat?cac?atc?cac?agg?att?ctc?aaa????528

Ile?Asn?Asp?Gly?Ile?Leu?Leu?Arg?Asn?His?Ile?His?Arg?Ile?Leu?Lys

165?????????????????170?????????????????175

aag?cat?ttc?cgt?gat?aag?cct?tac?tat?gtt?gac?ctt?gtt?gat?ttg?ttt????576

Lys?His?Phe?Arg?Asp?Lys?Pro?Tyr?Tyr?Val?Asp?Leu?Val?Asp?Leu?Phe

180?????????????????185?????????????????190

aat?gag?gtt?gag?ttg?caa?aca?gct?tgt?ggc?cag?atg?ata?gat?ttg?atc????624

Asn?Glu?Val?Glu?Leu?Gln?Thr?Ala?Cys?Gly?Gln?Met?Ile?Asp?Leu?Ile

195?????????????????200?????????????????205

acc?acc?ttt?gaa?gga?gaa?aag?gat?ttg?gcc?aag?tac?tca?ttg?tca?atc????672

Thr?Thr?Phe?Glu?Gly?Glu?Lys?Asp?Leu?Ala?Lys?Tyr?Ser?Leu?Ser?Ile

210?????????????????215?????????????????220

cac?cgt?cgt?att?gtc?cag?tac?aaa?acg?gct?tat?tac?tca?ttt?tat?ctc????720

His?Arg?Arg?Ile?Val?Gln?Tyr?Lys?Thr?Ala?Tyr?Tyr?Ser?Phe?Tyr?Leu

225?????????????????230?????????????????235?????????????????240

cct?gtt?gct?tgt?gcg?ttg?ctt?atg?gcg?ggc?gaa?aat?ttg?gaa?aac?cat????768

Pro?Val?Ala?Cys?Ala?Leu?Leu?Met?Ala?Gly?Glu?Asn?Leu?Glu?Asn?His

245?????????????????250?????????????????255

att?gac?gtg?aaa?aat?gtt?ctt?gtt?gac?atg?gga?atc?tac?ttc?caa?gtg????816

Ile?Asp?Val?Lys?Asn?Val?Leu?Val?Asp?Met?Gly?Ile?Tyr?Phe?Gln?Val

260?????????????????265?????????????????270

cag?gat?gat?tat?ctg?gat?tgt?ttt?gct?gat?ccc?gag?acg?ctt?ggc?aag?????864

Gln?Asp?Asp?Tyr?Leu?Asp?Cys?Phe?Ala?Asp?Pro?Glu?Thr?Leu?Gly?Lys

275?????????????????280?????????????????285

ata?gga?aca?gat?ata?gaa?gat?ttc?aaa?tgc?tcg?tgg?ttg?gtg?gtt?aag?????912

Ile?Gly?Thr?Asp?Ile?Glu?Asp?Phe?Lys?Cys?Ser?Trp?Leu?Val?Val?Lys

290?????????????????295?????????????????300

gca?tta?gag?cgc?tgc?agc?gaa?gaa?caa?act?aag?ata?tta?tat?gag?aac?????960

Ala?Leu?Glu?Arg?Cys?Ser?Glu?Glu?Gin?Thr?Lys?Ile?Leu?Tyr?Glu?Asn

305?????????????????310?????????????????315?????????????????320

tat?ggt?aaa?ccc?gac?cca?tcg?aac?gtt?gct?aaa?gtg?aag?gat?ctc?tac????1008

Tyr?Gly?Lys?Pro?Asp?Pro?Ser?Asn?Val?Ala?Lys?Val?Lys?Asp?Leu?Tyr

325?????????????????330?????????????????335

aaa?gag?ctg?gat?ctt?gag?gga?gtt?ttc?atg?gag?tat?gag?agc?aaa?agc????1056

Lys?Glu?Leu?Asp?Leu?Glu?Gly?Val?Phe?Met?Glu?Tyr?Glu?Ser?Lys?Ser

340?????????????????345?????????????????350

tac?gag?aag?ctg?act?gga?gcg?att?gag?gga?cac?caa?agt?aaa?gca?atc????1104

Tyr?Glu?Lys?Leu?Thr?Gly?Ala?Ile?Glu?Gly?His?Gln?Ser?Lys?Ala?Ile

355?????????????????360?????????????????365

caa?gca?gtg?cta?aaa?tcc?ttc?ttg?gct?aag?atc?tac?aag?agg?cag?aag????1152

Gln?Ala?Val?Leu?Lye?Ser?Phe?Leu?Ala?Lys?Ile?Tyr?Lys?Arg?Gln?Lys

370?????????????????375?????????????????380

tag????????????????????????????????????????????????????????????????1155

<210>124

<211>384

<212>PRT

＜213〉mouseearcress

<400>124

Met?Ser?Val?Ser?Cys?Cys?Cys?Arg?Asn?Leu?Gly?Lys?Thr?Ile?Lys?Lys

1???????????????5???????????????????10??????????????????15

Ala?Ile?Pro?Ser?His?His?Leu?His?Leu?Arg?Ser?Leu?Gly?Gly?Ser?Leu

20??????????????????25??????????????????30

Tyr?Arg?Arg?Arg?Ile?Gln?Ser?Ser?Ser?Met?Glu?Thr?Asp?Leu?Lys?Ser

35??????????????????40??????????????????45

Thr?Phe?Leu?Asn?Val?Tyr?Ser?Val?Leu?Lys?Ser?Asp?Leu?Leu?His?Asp

50??????????????????55??????????????????60

Pro?Ser?Phe?Glu?Phe?Thr?Asn?Glu?Ser?Arg?Leu?Trp?Val?Asp?Arg?Met

65??????????????????70??????????????????75??????????????????80

Leu?Asp?Tyr?Asn?Val?Arg?Gly?Gly?Lys?Leu?Asn?Arg?Gly?Leu?Ser?Val

85??????????????????90??????????????????95

Val?Asp?Ser?Phe?Lys?Leu?Leu?Lys?Gln?Gly?Asn?Asp?Leu?Thr?Glu?Gln

100?????????????????105?????????????????110

Glu?Val?Phe?Leu?Ser?Cys?Ala?Leu?Gly?Trp?Cys?Ile?Glu?Trp?Leu?Gln

115?????????????????120?????????????????125

Ala?Tyr?Phe?Leu?Val?Leu?Asp?Asp?Ile?Met?Asp?Asn?Ser?Val?Thr?Arg

130?????????????????135?????????????????140

Arg?Gly?Gln?Pro?Cys?Trp?Phe?Arg?Val?Pro?Gln?Val?Gly?Met?Val?Ala

145?????????????????150?????????????????155?????????????????160

Ile?Asn?Asp?Gly?Ile?Leu?Leu?Arg?Asn?His?Ile?His?Arg?Ile?Leu?Lys

165?????????????????170?????????????????175

Lys?His?Phe?Arg?Asp?Lys?Pro?Tyr?Tyr?Val?Asp?Leu?Val?Asp?Leu?Phe

180?????????????????185?????????????????190

Asn?Glu?Val?Glu?Leu?Gln?Thr?Ala?Cys?Gly?Gln?Met?Ile?Asp?Leu?Ile

195?????????????????200?????????????????205

Thr?Thr?Phe?Glu?Gly?Glu?Lys?Asp?Leu?Ala?Lys?Tyr?Ser?Leu?Ser?Ile

210?????????????????215?????????????????220

His?Arg?Arg?Ile?Val?Gln?Tyr?Lys?Thr?Ala?Tyr?Tyr?Ser?Phe?Tyr?Leu

225?????????????????230?????????????????235?????????????????240

Pro?Val?Ala?Cys?Ala?Leu?Leu?Met?Ala?Gly?Glu?Asn?Leu?Glu?Asn?His

245?????????????????250?????????????????255

Ile?Asp?Val?Lys?Asn?Val?Leu?Val?Asp?Met?Gly?Ile?Tyr?Phe?Gln?Val

260?????????????????265?????????????????270

Gln?Asp?Asp?Tyr?Leu?Asp?Cys?Phe?Ala?Asp?Pro?Glu?Thr?Leu?Gly?Lys

275?????????????????280?????????????????285

Ile?Gly?Thr?Asp?Ile?Glu?Asp?Phe?Lys?Cys?Ser?Trp?Leu?Val?Val?Lys

290?????????????????295?????????????????300

Ala?Leu?Glu?Arg?Cys?Ser?Glu?Glu?Gln?Thr?Lys?Ile?Leu?Tyr?Glu?Asn

305?????????????????310?????????????????315?????????????????320

Tyr?Gly?Lys?Pro?Asp?Pro?Ser?Asn?Val?Ala?Lys?Val?Lys?Asp?Leu?Tyr

325?????????????????330?????????????????335

Lys?Glu?Leu?Asp?Leu?Glu?Gly?Val?Phe?Met?Glu?Tyr?Glu?Ser?Lys?Ser

340?????????????????345?????????????????350

Tyr?Glu?Lys?Leu?Thr?Gly?Ala?Ile?Glu?Gly?His?Gln?Ser?Lys?Ala?Ile

355?????????????????360?????????????????365

Gln?Ala?Val?Leu?Lys?Ser?Phe?Leu?Ala?Lys?Ile?Tyr?Lys?Arg?Gln?Lys

370?????????????????375?????????????????380

<210>125

<211>1101

<212>DNA

＜213〉sinapsis alba (Sinabs alba)

<220>

<221>CDS

<222>(1)..(1101)

<223>

<400>125

atg?gct?tct?tca?gtg?act?cct?cta?ggt?tca?tgg?gtt?ctt?ctt?cac?cat????48

Met?Ala?Ser?Ser?Val?Thr?Pro?Leu?Gly?Ser?Trp?Val?Leu?Leu?His?His

1???????????????5???????????????????10??????????????????15

cat?cct?tca?act?atc?tta?acc?caa?tcc?aga?tcc?aga?tct?cct?cct?tct????96

His?Pro?Ser?Thr?Ile?Leu?Thr?Gln?Ser?Arg?Ser?Arg?Ser?Pro?Pro?Ser

20??????????????????25??????????????????30

ctc?atc?acc?ctt?aaa?ccc?atc?tcc?ctc?act?cca?aaa?cgc?acc?gtt?tcg????144

Leu?Ile?Thr?Leu?Lys?Pro?Ile?Ser?Leu?Thr?Pro?Lys?Arg?Thr?Val?Ser

35??????????????????40??????????????????45

tct?tct?tcc?tcc?tct?tcc?ctc?atc?acc?aaa?gaa?gac?aac?aac?ctc?aaa????192

Ser?Ser?Ser?Ser?Ser?Ser?Leu?Ile?Thr?Lys?Glu?Asp?Asn?Asn?Leu?Lys

50??????????????????55??????????????????60

tcc?tct?tcc?tct?tcc?ttc?gat?ttc?atg?tct?tac?atc?atc?cgc?aaa?gcc????240

Ser?Ser?Ser?Ser?Ser?Phe?Asp?Phe?Met?Ser?Tyr?Ile?Ile?Arg?Lys?Ala

65??????????????????70??????????????????75??????????????????80

gac?tcc?gtc?aac?aaa?gcc?tta?gac?tcc?gcc?gtc?cct?ctc?cgg?gag?cca????288

Asp?Ser?Val?Asn?Lys?Ala?Leu?Asp?Ser?Ala?Val?Pro?Leu?Arg?Glu?Pro

85??????????????????90??????????????????95

ctc?aag?atc?cac?gaa?gcg?atg?cgt?tac?tct?ctc?ctc?gcc?gga?gga?aaa????336

Leu?Lys?Ile?His?Glu?Ala?Met?Arg?Tyr?Ser?Leu?Leu?Ala?Gly?Gly?Lys

100?????????????????105?????????????????110

cgc?gtc?aga?cca?gtt?ctc?tgc?atc?gcc?gcg?tgc?gag?cta?gtc?gga?gga????384

Arg?Val?Arg?Pro?Val?Leu?Cys?Ile?Ala?Ala?Cys?Glu?Leu?Val?Gly?Gly

115?????????????????120?????????????????125

gaa?gag?tct?tta?gct?atg?ccg?gcg?cgt?tgc?gcc?gtg?gaa?atg?atc?cac????432

Glu?Glu?Ser?Leu?Ala?Met?Pro?Ala?Arg?Cys?Ala?Val?Glu?Met?Ile?His

130?????????????????135?????????????????140

acc?atg?tcg?ttg?atc?cac?gac?gac?ttg?cct?tgt?atg?gat?aac?gac?gat????480

Thr?Met?Ser?Leu?Ile?His?Asp?Asp?Leu?Pro?Cys?Met?Asp?Asn?Asp?Asp

145?????????????????150?????????????????155?????????????????160

ctc?cgc?cgc?gga?aag?ccc?acg?aat?cac?aaa?gtt?tac?ggc?gaa?gac?gtg????528

Leu?Arg?Arg?Gly?Lys?Pro?Thr?Asn?His?Lys?Val?Tyr?Gly?Glu?Asp?Val

165?????????????????170?????????????????175

gcg?gtt?tta?gcc?gga?gac?gcg?ctt?ctt?tcg?ttc?gcc?ttc?gag?cat?tta????576

Ala?Val?Leu?Ala?Gly?Asp?Ala?Leu?Leu?Ser?Phe?Ala?Phe?Glu?His?Leu

180?????????????????185?????????????????190

gcg?tcg?gct?acg?agc?tcg?gag?gtt?tct?ccg?gcg?aga?gtg?gtt?aga?gct????624

Ala?Ser?Ala?Thr?Ser?Ser?Glu?Val?Ser?Pro?Ala?Arg?Val?Val?Arg?Ala

195?????????????????200?????????????????205

gtg?gga?gag?ttg?gct?aaa?gcc?atc?ggc?acc?gaa?ggg?ctc?gtg?gcg?gga????672

Val?Gly?Glu?Leu?Ala?Lys?Ala?Ile?Gly?Thr?Glu?Gly?Leu?Val?Ala?Gly

210?????????????????215?????????????????220

caa?gtg?gtg?gat?ata?agc?agt?gaa?ggg?ttg?gac?tta?aac?aac?gtc?gga????720

Gln?Val?Val?Asp?Ile?Ser?Ser?Glu?Gly?Leu?Asp?Leu?Asn?Asn?Val?Gly

225?????????????????230?????????????????235?????????????????240

ttg?gag?cat?ttg?aag?ttt?ata?cat?ttg?cat?aaa?acg?gcg?gcg?ttg?ctt????768

Leu?Glu?His?Leu?Lys?Phe?Ile?His?Leu?His?Lys?Thr?Ala?Ala?Leu?Leu

245?????????????????250?????????????????255

gaa?gct?tca?gcg?gtt?ttg?ggt?ggg?atc?atc?ggt?gga?ggg?agt?gat?gaa????816

Glu?Ala?Ser?Ala?Val?Leu?Gly?Gly?Ile?Ile?Gly?Gly?Gly?Ser?Asp?Glu

260?????????????????265?????????????????270

gag?atc?gag?agg?ctg?agg?aag?ttc?gcg?agg?tgt?att?ggg?ttg?ttg?ttt????864

Glu?Ile?Glu?Arg?Leu?Arg?Lys?Phe?Ala?Arg?Cys?Ile?Gly?Leu?Leu?Phe

275?????????????????280?????????????????285

cag?gtg?gtt?gat?gat?atc?ttg?gac?gtg?acg?aaa?tcg?tct?caa?gaa?ctg????912

Gln?Val?Val?Asp?Asp?Ile?Leu?Asp?Val?Thr?Lys?Ser?Ser?Gln?Glu?Leu

290?????????????????295?????????????????300

ggg?aaa?acc?gct?ggg?aaa?gat?ttg?att?gct?gat?aag?ttg?act?tat?ccg????960

Gly?Lys?Thr?Ala?Gly?Lys?Asp?Leu?Ile?Ala?Asp?Lys?Leu?Thr?Tyr?Pro

305?????????????????310?????????????????315?????????????????320

aag?ctc?atg?ggt?ttg?gag?aaa?tcg?aga?gag?ttc?gct?gag?aag?ttg?aat????1008

Lys?Leu?Met?Gly?Leu?Glu?Lys?Ser?Arg?Glu?Phe?Ala?Glu?Lys?Leu?Asn

325?????????????????330?????????????????335

aca?gag?gca?cgt?gat?cag?ctt?tta?ggg?ttt?gat?tcc?gac?aag?gtt?gct????1056

Thr?Glu?Ala?Arg?Asp?Gln?Leu?Leu?Gly?Phe?Asp?Ser?Asp?Lys?Val?Ala

340?????????????????345?????????????????350

cct?ttg?ttg?gct?ttg?gct?aat?tac?att?gcc?aat?aga?cag?aac?tga????????1101

Pro?Leu?Leu?Ala?Leu?Ala?Asn?Tyr?Ile?Ala?Asn?Arg?Gln?Asn

355?????????????????360?????????????????365

<210>126

<211>366

<212>PRT

＜213〉sinapsis alba

<400>126

Met?Ala?Ser?Ser?Val?Thr?Pro?Leu?Gly?Ser?Trp?Val?Leu?Leu?His?His

1???????????????5???????????????????10??????????????????15

His?Pro?Ser?Thr?Ile?Leu?Thr?Gln?Ser?Arg?Ser?Arg?Ser?Pro?Pro?Ser

20??????????????????25??????????????????30

Leu?Ile?Thr?Leu?Lys?Pro?Ile?Ser?Leu?Thr?Pro?Lys?Arg?Thr?Val?Ser

35??????????????????40??????????????????45

Ser?Ser?Ser?Ser?Ser?Ser?Leu?Ile?Thr?Lys?Glu?Asp?Asn?Asn?Leu?Lys

50??????????????????55??????????????????60

Ser?Ser?Ser?Ser?Ser?Phe?Asp?Phe?Met?Ser?Tyr?Ile?Ile?Arg?Lys?Ala

65??????????????????70??????????????????75??????????????????80

Asp?Ser?Val?Asn?Lys?Ala?Leu?Asp?Ser?Ala?Val?Pro?Leu?Arg?Glu?Pro

85??????????????????90??????????????????95

Leu?Lys?Ile?His?Glu?Ala?Met?Arg?Tyr?Ser?Leu?Leu?Ala?Gly?Gly?Lys

100?????????????????105?????????????????110

Arg?Val?Arg?Pro?Val?Leu?Cys?Ile?Ala?Ala?Cys?Glu?Leu?Val?Gly?Gly

115?????????????????120?????????????????125

Glu?Glu?Ser?Leu?Ala?Met?Pro?Ala?Arg?Cys?Ala?Val?Glu?Met?Ile?His

130?????????????????135?????????????????140

Thr?Met?Ser?Leu?Ile?His?Asp?Asp?Leu?Pro?Cys?Met?Asp?Asn?Asp?Asp

145?????????????????150?????????????????155?????????????????160

Leu?Arg?Arg?Gly?Lys?Pro?Thr?Asn?His?Lys?Val?Tyr?Gly?Glu?Asp?Val

165?????????????????170?????????????????175

Ala?Val?Leu?Ala?Gly?Asp?Ala?Leu?Leu?Ser?Phe?Ala?Phe?Glu?His?Leu

180?????????????????185?????????????????190

Ala?Ser?Ala?Thr?Ser?Ser?Glu?Val?Ser?Pro?Ala?Arg?Val?Val?Arg?Ala

195?????????????????200?????????????????205

Val?Gly?Glu?Leu?Ala?Lys?Ala?Ile?Gly?Thr?Glu?Gly?Leu?Val?Ala?Gly

210?????????????????215?????????????????220

Gln?Val?Val?Asp?Ile?Ser?Ser?Glu?Gly?Leu?Asp?Leu?Asn?Asn?Val?Gly

225?????????????????230?????????????????235?????????????????240

Leu?Glu?His?Leu?Lys?Phe?Ile?His?Leu?His?Lys?Thr?Ala?Ala?Leu?Leu

245?????????????????250?????????????????255

Glu?Ala?Ser?Ala?Val?Leu?Gly?Gly?Ile?Ile?Gly?Gly?Gly?Ser?Asp?Glu

260?????????????????265?????????????????270

Glu?Ile?Glu?Arg?Leu?Arg?Lys?Phe?Ala?Arg?Cys?Ile?Gly?Leu?Leu?Phe

275?????????????????280?????????????????285

Gln?Val?Val?Asp?Asp?Ile?Leu?Asp?Val?Thr?Lys?Ser?Ser?Gln?Glu?Leu

290?????????????????295?????????????????300

Gly?Lys?Thr?Ala?Gly?Lys?Asp?Leu?Ile?Ala?Asp?Lys?Leu?Thr?Tyr?Pro

305?????????????????310?????????????????315?????????????????320

Lys?Leu?Met?Gly?Leu?Glu?Lys?Ser?Arg?Glu?Phe?Ala?Glu?Lys?Leu?Asn

325?????????????????330?????????????????335

Thr?Glu?Ala?Arg?Asp?Gln?Leu?Leu?Gly?Phe?Asp?Ser?Asp?Lys?Val?Ala

340?????????????????345?????????????????350

Pro?Leu?Leu?Ala?Leu?Ala?Asn?Tyr?Ile?Ala?Asn?Arg?Gln?Asn

355?????????????????360?????????????????365

<210>127

<211>930

<212>DNA

＜213〉Erwinia uredovora (Erwinia uredovora)

<220>

<221>CDS

<222>(1)..(930)

<223>

<400>127

atg?aat?aat?ccg?tcg?tta?ctc?aat?cat?gcg?gtc?gaa?acg?atg?gca?gtt????48

Met?Asn?Asn?Pro?Ser?Leu?Leu?Asn?His?Ala?Val?Glu?Thr?Met?Ala?Val

1???????????????5???????????????????10??????????????????15

ggc?tcg?aaa?agt?ttt?gcg?aca?gcc?tca?aag?tta?ttt?gat?gca?aaa?acc????96

Gly?Ser?Lys?Ser?Phe?Ala?Thr?Ala?Ser?Lys?Leu?Phe?Asp?Ala?Lys?Thr

20??????????????????25??????????????????30

cgg?cgc?agc?gta?ctg?atg?ctc?tac?gcc?tgg?tgc?cgc?cat?tgt?gac?gat????144

Arg?Arg?Ser?Val?Leu?Met?Leu?Tyr?Ala?Trp?Cys?Arg?His?Cys?Asp?Asp

35??????????????????40??????????????????45

gtt?att?gac?gat?cag?acg?ctg?ggc?ttt?cag?gcc?cgg?cag?cct?gcc?tta????192

Val?Ile?Asp?Asp?Gln?Thr?Leu?Gly?Phe?Gln?Ala?Arg?Gln?Pro?Ala?Leu

50??????????????????55??????????????????60

caa?acg?ccc?gaa?caa?cgt?ctg?atg?caa?ctt?gag?atg?aaa?acg?cgc?cag????240

Gln?Thr?Pro?Glu?Gln?Arg?Leu?Met?Gln?Leu?Glu?Met?Lys?Thr?Arg?Gln

65??????????????????70??????????????????75??????????????????80

gcc?tat?gca?gga?tcg?cag?atg?cac?gaa?ccg?gcg?ttt?gcg?gct?ttt?cag????288

Ala?Tyr?Ala?Gly?Ser?Gln?Met?His?Glu?Pro?Ala?Phe?Ala?Ala?Phe?Gln

85??????????????????90??????????????????95

gaa?gtg?gct?atg?gct?cat?gat?atc?gcc?ccg?gct?tac?gcg?ttt?gat?cat????336

Glu?Val?Ala?Met?Ala?His?Asp?Ile?Ala?Pro?Ala?Tyr?Ala?Phe?Asp?His

100?????????????????105?????????????????110

ctg?gaa?ggc?ttc?gcc?atg?gat?gta?cgc?gaa?gcg?caa?tac?agc?caa?ctg????384

Leu?Glu?Gly?Phe?Ala?Met?Asp?Val?Arg?Glu?Ala?Gln?Tyr?Ser?Gln?Leu

115?????????????????120?????????????????125

gat?gat?acg?ctg?cgc?tat?tgc?tat?cac?gtt?gca?ggc?gtt?gtc?ggc?ttg????432

Asp?Asp?Thr?Leu?Arg?Tyr?Cys?Tyr?His?Val?Ala?Gly?Val?Val?Gly?Leu

130?????????????????135?????????????????140

atg?atg?gcg?caa?atc?atg?ggc?gtg?cgg?gat?aac?gcc?acg?ctg?gac?cgc????480

Met?Met?Ala?Gln?Ile?Met?Gly?Val?Arg?Asp?Asn?Ala?Thr?Leu?Asp?Arg

145?????????????????150?????????????????155?????????????????160

gcc?tgt?gac?ctt?ggg?ctg?gca?ttt?cag?ttg?acc?aat?att?gct?cgc?gat????528

Ala?Cys?Asp?Leu?Gly?Leu?Ala?Phe?Gln?Leu?Thr?Asn?Ile?Ala?Arg?Asp

165?????????????????170?????????????????175

att?gtg?gac?gat?gcg?cat?gcg?ggc?cgc?tgt?tat?ctg?ccg?gca?agc?tgg????576

Ile?Val?Asp?Asp?Ala?His?Ala?Gly?Arg?Cys?Tyr?Leu?Pro?Ala?Ser?Trp

180?????????????????185?????????????????190

ctg?gag?cat?gaa?ggt?ctg?aac?aaa?gag?aat?tat?gcg?gca?cct?gaa?aac????624

Leu?Glu?His?Glu?Gly?Leu?Asn?Lys?Glu?Asn?Tyr?Ala?Ala?Pro?Glu?Asn

195?????????????????200?????????????????205

cgt?cag?gcg?ctg?agc?cgt?atc?gcc?cgt?cgt?ttg?gtg?cag?gaa?gca?gaa????672

Arg?Gln?Ala?Leu?Ser?Arg?Ile?Ala?Arg?Arg?Leu?Val?Gln?Glu?Ala?Glu

210?????????????????215?????????????????220

cct?tac?tat?ttg?tct?gcc?aca?gcc?ggc?ctg?gca?ggg?ttg?ccc?ctg?cgt????720

Pro?Tyr?Tyr?Leu?Ser?Ala?Thr?Ala?Gly?Leu?Ala?Gly?Leu?Pro?Leu?Arg

225?????????????????230?????????????????235?????????????????240

tcc?gcc?tgg?gca?atc?gct?acg?gcg?aag?cag?gtt?tac?cgg?aaa?ata?ggt????768

Ser?Ala?Trp?Ala?Ile?Ala?Thr?Ala?Lys?Gln?Val?Tyr?Arg?Lys?Ile?Gly

245?????????????????250?????????????????255

gtc?aaa?gtt?gaa?cag?gcc?ggt?cag?caa?gcc?tgg?gat?cag?cgg?cag?tca????816

Val?Lys?Val?Glu?Gln?Ala?Gly?Gln?Gln?Ala?Trp?Asp?Gln?Arg?Gln?Ser

260?????????????????265?????????????????270

acg?acc?acg?ccc?gaa?aaa?tta?acg?ctg?ctg?ctg?gcc?gcc?tct?ggt?cag????864

Thr?Thr?Thr?Pro?Glu?Lys?Leu?Thr?Leu?Leu?Leu?Ala?Ala?Ser?Gly?Gln

275?????????????????280?????????????????285

gcc?ctt?act?tcc?cgg?atg?cgg?gct?cat?cct?ccc?cgc?cct?gcg?cat?ctc????912

Ala?Leu?Thr?Ser?Arg?Met?Arg?Ala?His?Pro?Pro?Arg?Pro?Ala?His?Leu

290?????????????????295?????????????????300

tgg?cag?cgc?ccg?ctc?tag????????????????????????????????????????????930

Trp?Gln?Arg?Pro?Leu

305

<210>128

<211>309

<212>PRT

＜213〉Erwinia uredovora

<400>128

Met?Asn?Asn?Pro?Ser?Leu?Leu?Asn?His?Ala?Val?Glu?Thr?Met?Ala?Val

1???????????????5???????????????????10??????????????????15

Gly?Ser?Lys?Ser?Phe?Ala?Thr?Ala?Ser?Lys?Leu?Phe?Asp?Ala?Lys?Thr

20??????????????????25??????????????????30

Arg?Arg?Ser?Val?Leu?Met?Leu?Tyr?Ala?Trp?Cys?Arg?His?Cys?Asp?Asp

35??????????????????40??????????????????45

Val?Ile?Asp?Asp?Gln?Thr?Leu?Gly?Phe?Gln?Ala?Arg?Gln?Pro?Ala?Leu

50??????????????????55??????????????????60

Gln?Thr?Pro?Glu?Gln?Arg?Leu?Met?Gln?Leu?Glu?Met?Lys?Thr?Arg?Gln

65??????????????????70??????????????????75??????????????????80

Ala?Tyr?Ala?Gly?Ser?Gln?Met?His?Glu?Pro?Ala?Phe?Ala?Ala?Phe?Gln

85??????????????????90??????????????????95

Glu?Val?Ala?Met?Ala?His?Asp?Ile?Ala?Pro?Ala?Tyr?Ala?Phe?Asp?His

100?????????????????105?????????????????110

Leu?Glu?Gly?Phe?Ala?Met?Asp?Val?Arg?Glu?Ala?Gln?Tyr?Ser?Gln?Leu

115?????????????????120?????????????????125

Asp?Asp?Thr?Leu?Arg?Tyr?Cys?Tyr?His?Val?Ala?Gly?Val?Val?Gly?Leu

130?????????????????135?????????????????140

Met?Met?Ala?Gln?Ile?Met?Gly?Val?Arg?Asp?Asn?Ala?Thr?Leu?Asp?Arg

145?????????????????150?????????????????155?????????????????160

Ala?Cys?Asp?Leu?Gly?Leu?Ala?Phe?Gln?Leu?Thr?Asn?Ile?Ala?Arg?Asp

165?????????????????170?????????????????175

Ile?Val?Asp?Asp?Ala?His?Ala?Gly?Arg?Cys?Tyr?Leu?Pro?Ala?Ser?Trp

180?????????????????185?????????????????190

Leu?Glu?His?Glu?Gly?Leu?Asn?Lys?Glu?Asn?Tyr?Ala?Ala?Pro?Glu?Asn

195?????????????????200?????????????????205

Arg?Gln?Ala?Leu?Ser?Arg?Ile?Ala?Arg?Arg?Leu?Val?Gln?Glu?Ala?Glu

210?????????????????215?????????????????220

Pro?Tyr?Tyr?Leu?Ser?Ala?Thr?Ala?Gly?Leu?Ala?Gly?Leu?Pro?Leu?Arg

225?????????????????230?????????????????235?????????????????240

Ser?Ala?Trp?Ala?Ile?Ala?Thr?Ala?Lys?Gln?Val?Tyr?Arg?Lys?Ile?Gly

245?????????????????250?????????????????255

Val?Lys?Val?Glu?Gln?Ala?Gly?Gln?Gln?Ala?Trp?Asp?Gln?Arg?Gln?Ser

260?????????????????265?????????????????270

Thr?Thr?Thr?Pro?Glu?Lys?Leu?Thr?Leu?Leu?Leu?Ala?Ala?Ser?Gly?Gln

275?????????????????280?????????????????285

Ala?Leu?Thr?Ser?Arg?Met?Arg?Ala?His?Pro?Pro?Arg?Pro?Ala?His?Leu

290?????????????????295?????????????????300

Trp?Gln?Arg?Pro?Leu

305

<210>129

<211>1479

<212>DNA

＜213〉Erwinia uredovora

<220>

<221>CDS

<222>(1)..(1479)

<223>

<400>129

atg?aaa?cca?act?acg?gta?att?ggt?gca?ggc?ttc?ggt?ggc?ctg?gca?ctg????48

Met?Lys?Pro?Thr?Thr?Val?Ile?Gly?Ala?Gly?Phe?Gly?Gly?Leu?Ala?Leu

1???????????????5???????????????????10??????????????????15

gca?att?cgt?cta?caa?gct?gcg?ggg?atc?ccc?gtc?tta?ctg?ctt?gaa?caa????96

Ala?Ile?Arg?Leu?Gln?Ala?Ala?Gly?Ile?Pro?Val?Leu?Leu?Leu?Glu?Gln

20??????????????????25??????????????????30

cgt?gat?aaa?ccc?ggc?ggt?cgg?gct?tat?gtc?tac?gag?gat?cag?ggg?ttt????144

Arg?Asp?Lys?Pro?Gly?Gly?Arg?Ala?Tyr?Val?Tyr?Glu?Asp?Gln?Gly?Phe

35??????????????????40??????????????????45

acc?ttt?gat?gca?ggc?ccg?acg?gtt?atc?acc?gat?ccc?agt?gcc?att?gaa????192

Thr?Phe?Asp?Ala?Gly?Pro?Thr?Val?Ile?Thr?Asp?Pro?Ser?Ala?Ile?Glu

50??????????????????55??????????????????60

gaa?ctg?ttt?gca?ctg?gca?gga?aaa?cag?tta?aaa?gag?tat?gtc?gaa?ctg????240

Glu?Leu?Phe?Ala?Leu?Ala?Gly?Lys?Gln?Leu?Lys?Glu?Tyr?Val?Glu?Leu

65??????????????????70??????????????????75??????????????????80

ctg?ccg?gtt?acg?ccg?ttt?tac?cgc?ctg?tgt?tgg?gag?tca?ggg?aag?gtc????288

Leu?Pro?Val?Thr?Pro?Phe?Tyr?Arg?Leu?Cys?Trp?Glu?Ser?Gly?Lys?Val

85??????????????????90??????????????????95

ttt?aat?tac?gat?aac?gat?caa?acc?cgg?ctc?gaa?gcg?cag?att?cag?cag????336

Phe?Asn?Tyr?Asp?Asn?Asp?Gln?Thr?Arg?Leu?Glu?Ala?Gln?Ile?Gln?Gln

100?????????????????105?????????????????110

ttt?aat?ccc?cgc?gat?gtc?gaa?ggt?tat?cgt?cag?ttt?ctg?gac?tat?tca????384

Phe?Asn?Pro?Arg?Asp?Val?Glu?Gly?Tyr?Arg?Gln?Phe?Leu?Asp?Tyr?Ser

115?????????????????120?????????????????125

cgc?gcg?gtg?ttt?aaa?gaa?ggc?tat?cta?aag?ctc?ggt?act?gtc?cct?ttt????432

Arg?Ala?Val?Phe?Lys?Glu?Gly?Tyr?Leu?Lys?Leu?Gly?Thr?Val?Pro?Phe

130?????????????????135?????????????????140

tta?tcg?ttc?aga?gac?atg?ctt?cgc?gcc?gca?cct?caa?ctg?gcg?aaa?ctg????480

Leu?Ser?Phe?Arg?Asp?Met?Leu?Arg?Ala?Ala?Pro?Gln?Leu?Ala?Lys?Leu

145?????????????????150?????????????????155?????????????????160

cag?gca?tgg?aga?agc?gtt?tac?agt?aag?gtt?gcc?agt?tac?atc?gaa?gat????528

Gln?Ala?Trp?Arg?Ser?Val?Tyr?Ser?Lys?Val?Ala?Ser?Tyr?Ile?Glu?Asp

165?????????????????170?????????????????175

gaa?cat?ctg?cgc?cag?gcg?ttt?tct?ttc?cac?tcg?ctg?ttg?gtg?ggc?ggc????576

Glu?His?Leu?Arg?Gln?Ala?Phe?Ser?Phe?His?Ser?Leu?Leu?Val?Gly?Gly

180?????????????????185?????????????????190

aat?ccc?ttc?gcc?acc?tca?tcc?att?tat?acg?ttg?ata?cac?gcg?ctg?gag????624

Asn?Pro?Phe?Ala?Thr?Ser?Ser?Ile?Tyr?Thr?Leu?Ile?His?Ala?Leu?Glu

195?????????????????200?????????????????205

cgt?gag?tgg?ggc?gtc?tgg?ttt?ccg?cgt?ggc?ggc?acc?ggc?gca?tta?gtt????672

Arg?Glu?Trp?Gly?Val?Trp?Phe?Pro?Arg?Gly?Gly?Thr?Gly?Ala?Leu?Val

210?????????????????215?????????????????220

cag?ggg?atg?ata?aag?ctg?ttt?cag?gat?ctg?ggt?ggc?gaa?gtc?gtg?tta????720

Gln?Gly?Met?Ile?Lys?Leu?Phe?Gln?Asp?Leu?Gly?Gly?Glu?Val?Val?Leu

225?????????????????230?????????????????235?????????????????240

aac?gcc?aga?gtc?agc?cat?atg?gaa?acg?aca?gga?aac?aag?att?gaa?gcc????768

Asn?Ala?Arg?Val?Ser?His?Met?Glu?Thr?Thr?Gly?Asn?Lys?Ile?Glu?Ala

245?????????????????250?????????????????255

gtg?cat?tta?gag?gac?ggt?cgc?agg?ttc?ctg?acg?caa?gcc?gtc?gcg?tca????816

Val?His?Leu?Glu?Asp?Gly?Arg?Arg?Phe?Leu?Thr?Gln?Ala?Val?Ala?Ser

260?????????????????265?????????????????270

aat?gca?gat?gtg?gtt?cat?acc?tat?cgc?gac?ctg?tta?agc?cag?cac?cct????864

Asn?Ala?Asp?Val?Val?His?Thr?Tyr?Arg?Asp?Leu?Leu?Ser?Gln?His?Pro

275?????????????????280?????????????????285

gcc?gcg?gtt?aag?cag?tcc?aac?aaa?ctg?cag?act?aag?cgc?atg?agt?aac????912

Ala?Ala?Val?Lys?Gln?Ser?Asn?Lys?Leu?Gln?Thr?Lys?Arg?Met?Ser?Asn

290?????????????????295?????????????????300

tct?ctg?ttt?gtg?ctc?tat?ttt?ggt?ttg?aat?cac?cat?cat?gat?cag?ctc????960

Ser?Leu?Phe?Val?Leu?Tyr?Phe?Gly?Leu?Asn?His?His?His?Asp?Gln?Leu

305?????????????????310?????????????????315?????????????????320

gcg?cat?cac?acg?gtt?tgt?ttc?ggc?ccg?cgt?tac?cgc?gag?ctg?att?gac????1008

Ala?His?His?Thr?Val?Cys?Phe?Gly?Pro?Arg?Tyr?Arg?Glu?Leu?Ile?Asp

325?????????????????330?????????????????335

gaa?att?ttt?aat?cat?gat?ggc?ctc?gca?gag?gac?ttc?tca?ctt?tat?ctg????1056

Glu?Ile?Phe?Asn?His?Asp?Gly?Leu?Ala?Glu?Asp?Phe?Ser?Leu?Tyr?Leu

340?????????????????345?????????????????350

cac?gcg?ccc?tgt?gtc?acg?gat?tcg?tca?ctg?gcg?cct?gaa?ggt?tgc?ggc????1104

His?Ala?Pro?Cys?Val?Thr?Asp?Ser?Ser?Leu?Ala?Pro?Glu?Gly?Cys?Gly

355?????????????????360?????????????????365

agt?tac?tat?gtg?ttg?gcg?ccg?gtg?ccg?cat?tta?ggc?acc?gcg?aac?ctc????1152

Ser?Tyr?Tyr?Val?Leu?Ala?Pro?Val?Pro?His?Leu?Gly?Thr?Ala?Asn?Leu

370?????????????????375?????????????????380

gac?tgg?acg?gtt?gag?ggg?cca?aaa?cta?cgc?gac?cgt?att?ttt?gcg?tac????1200

Asp?Trp?Thr?Val?Glu?Gly?Pro?Lys?Leu?Arg?Asp?Arg?Ile?Phe?Ala?Tyr

385?????????????????390?????????????????395?????????????????400

ctt?gag?cag?cat?tac?atg?cct?ggc?tta?cgg?agt?cag?ctg?gtc?acg?cac????1248

Leu?Glu?Gln?His?Tyr?Met?Pro?Gly?Leu?Arg?Ser?Gln?Leu?Val?Thr?His

405?????????????????410?????????????????415

cgg?atg?ttt?acg?ccg?ttt?gat?ttt?cgc?gac?cag?ctt?aat?gcc?tat?cat????1296

Arg?Met?Phe?Thr?Pro?Phe?Asp?Phe?Arg?Asp?Gln?Leu?Asn?Ala?Tyr?His

420?????????????????425?????????????????430

ggc?tca?gcc?ttt?tct?gtg?gag?ccc?gtt?ctt?acc?cag?agc?gcc?tgg?ttt????1344

Gly?Ser?Ala?Phe?Ser?Val?Glu?Pro?Val?Leu?Thr?Gln?Ser?Ala?Trp?Phe

435?????????????????440?????????????????445

cgg?ccg?cat?aac?cgc?gat?aaa?acc?att?act?aat?ctc?tac?ctg?gtc?ggc????1392

Arg?Pro?His?Asn?Arg?Asp?Lys?Thr?Ile?Thr?Asn?Leu?Tyr?Leu?Val?Gly

450?????????????????455?????????????????460

gca?ggc?acg?cat?ccc?ggc?gca?ggc?att?cct?ggc?gtc?atc?ggc?tcg?gca????1440

Ala?Gly?Thr?His?Pro?Gly?Ala?Gly?Ile?Pro?Gly?Val?Ile?Gly?Ser?Ala

465?????????????????470?????????????????475?????????????????480

aaa?gcg?aca?gca?ggt?ttg?atg?ctg?gag?gat?ctg?ata?tga????????????????1479

Lys?Ala?Thr?Ala?Gly?Leu?Met?Leu?Glu?Asp?Leu?Ile

485?????????????????490

<210>130

<211>492

<212>PRT

＜213〉Erwinia uredovora

<400>130

Met?Lys?Pro?Thr?Thr?Val?Ile?Gly?Ala?Gly?Phe?Gly?Gly?Leu?Ala?Leu

1???????????????5???????????????????10??????????????????15

Ala?Ile?Arg?Leu?Gln?Ala?Ala?Gly?Ile?Pro?Val?Leu?Leu?Leu?Glu?Gln

20??????????????????25??????????????????30

Arg?Asp?Lys?Pro?Gly?Gly?Arg?Ala?Tyr?Val?Tyr?Glu?Asp?Gln?Gly?Phe

35??????????????????40??????????????????45

Thr?Phe?Asp?Ala?Gly?Pro?Thr?Val?Ile?Thr?Asp?Pro?Ser?Ala?Ile?Glu

50??????????????????55??????????????????60

Glu?Leu?Phe?Ala?Leu?Ala?Gly?Lys?Gln?Leu?Lys?Glu?Tyr?Val?Glu?Leu

65??????????????????70??????????????????75??????????????????80

Leu?Pro?Val?Thr?Pro?Phe?Tyr?Arg?Leu?Cys?Trp?Glu?Ser?Gly?Lys?Val

85??????????????????90??????????????????95

Phe?Asn?Tyr?Asp?Asn?Asp?Gln?Thr?Arg?Leu?Glu?Ala?Gln?Ile?Gln?Gln

100?????????????????105?????????????????110

Phe?Asn?Pro?Arg?Asp?Val?Glu?Gly?Tyr?Arg?Gln?Phe?Leu?Asp?Tyr?Ser

115?????????????????120?????????????????125

Arg?Ala?Val?Phe?Lys?Glu?Gly?Tyr?Leu?Lys?Leu?Gly?Thr?Val?Pro?Phe

130?????????????????135?????????????????140

Leu?Ser?Phe?Arg?Asp?Met?Leu?Arg?Ala?Ala?Pro?Gln?Leu?Ala?Lys?Leu

145?????????????????150?????????????????155?????????????????160

Gln?Ala?Trp?Arg?Ser?Val?Tyr?Ser?Lys?Val?Ala?Ser?Tyr?Ile?Glu?Asp

165?????????????????170?????????????????175

Glu?His?Leu?Arg?Gln?Ala?Phe?Ser?Phe?His?Ser?Leu?Leu?Val?Gly?Gly

180?????????????????185?????????????????190

Asn?Pro?Phe?Ala?Thr?Ser?Ser?Ile?Tyr?Thr?Leu?Ile?His?Ala?Leu?Glu

195?????????????????200?????????????????205

Arg?Glu?Trp?Gly?Val?Trp?Phe?Pro?Arg?Gly?Gly?Thr?Gly?Ala?Leu?Val

210?????????????????215?????????????????220

Gln?Gly?Met?Ile?Lys?Leu?Phe?Gln?Asp?Leu?Gly?Gly?Glu?Val?Val?Leu

225?????????????????230?????????????????235?????????????????240

Asn?Ala?Arg?Val?Ser?His?Met?Glu?Thr?Thr?Gly?Asn?Lys?Ile?Glu?Ala

245?????????????????250?????????????????255

Val?His?Leu?Glu?Asp?Gly?Arg?Arg?Phe?Leu?Thr?Gln?Ala?Val?Ala?Ser

260?????????????????265?????????????????270

Asn?Ala?Asp?Val?Val?His?Thr?Tyr?Arg?Asp?Leu?Leu?Ser?Gln?His?Pro

275?????????????????280?????????????????285

Ala?Ala?Val?Lys?Gln?Ser?Asn?Lys?Leu?Gln?Thr?Lys?Arg?Met?Ser?Asn

290?????????????????295?????????????????300

Ser?Leu?Phe?Val?Leu?Tyr?Phe?Gly?Leu?Asn?His?His?His?Asp?Gln?Leu

305?????????????????310?????????????????315?????????????????320

Ala?His?His?Thr?Val?Cys?Phe?Gly?Pro?Arg?Tyr?Arg?Glu?Leu?Ile?Asp

325?????????????????330?????????????????335

Glu?Ile?Phe?Asn?His?Asp?Gly?Leu?Ala?Glu?Asp?Phe?Ser?Leu?Tyr?Leu

340?????????????????345?????????????????350

His?Ala?Pro?Cys?Val?Thr?Asp?Ser?Ser?Leu?Ala?Pro?Glu?Gly?Cys?Gly

355?????????????????360?????????????????365

Ser?Tyr?Tyr?Val?Leu?Ala?Pro?Val?Pro?His?Leu?Gly?Thr?Ala?Asn?Leu

370?????????????????375?????????????????380

Asp?Trp?Thr?Val?Glu?Gly?Pro?Lys?Leu?Arg?Asp?Arg?Ile?Phe?Ala?Tyr

385?????????????????390?????????????????395?????????????????400

Leu?Glu?Gln?His?Tyr?Met?Pro?Gly?Leu?Arg?Ser?Gln?Leu?Val?Thr?His

405?????????????????410?????????????????415

Arg?Met?Phe?Thr?Pro?Phe?Asp?Phe?Arg?Asp?Gln?Leu?Asn?Ala?Tyr?His

420?????????????????425?????????????????430

Gly?Ser?Ala?Phe?Ser?Val?Glu?Pro?Val?Leu?Thr?Gln?Ser?Ala?Trp?Phe

435?????????????????440?????????????????445

Arg?Pro?His?Asn?Arg?Asp?Lys?Thr?Ile?Thr?Asn?Leu?Tyr?Leu?Val?Gly

450?????????????????455?????????????????460

Ala?Gly?Thr?His?Pro?Gly?Ala?Gly?Ile?Pro?Gly?Val?Ile?Gly?Ser?Ala

465?????????????????470?????????????????475?????????????????480

Lys?Ala?Thr?Ala?Gly?Leu?Met?Leu?Glu?Asp?Leu?Ile

485?????????????????490

<210>131

<211>1725

<212>DNA

＜213〉daffodil (Narcissus pseudonarcissus)

<220>

<221>CDS

<222>(1)..(1725)

<223>

<400>131

atg?gct?tct?tcc?act?tgt?tta?att?cat?tct?tcc?tct?ttt?ggg?gtt?gga????48

Met?Ala?Ser?Ser?Thr?Cys?Leu?Ile?His?Ser?Ser?Ser?Phe?Gly?Val?Gly

1???????????????5???????????????????10??????????????????15

gga?aag?aaa?gtg?aag?atg?aac?acg?atg?att?cga?tcg?aag?ttg?ttt?tca????96

Gly?Lys?Lys?Val?Lys?Met?Asn?Thr?Met?Ile?Arg?Ser?Lys?Leu?Phe?Ser

20??????????????????25??????????????????30

att?cgg?tcg?gct?ttg?gac?act?aag?gtg?tct?gat?atg?agc?gtc?aat?gct????144

Ile?Arg?Ser?Ala?Leu?Asp?Thr?Lys?Val?Ser?Asp?Met?Ser?Val?Asn?Ala

35??????????????????40??????????????????45

cca?aaa?gga?ttg?ttt?cca?cca?gag?cct?gag?cac?tac?agg?ggg?cca?aag????192

Pro?Lys?Gly?Leu?Phe?Pro?Pro?Glu?Pro?Glu?His?Tyr?Arg?Gly?Pro?Lys

50??????????????????55??????????????????60

ctt?aaa?gtg?gct?atc?att?gga?gct?ggg?ctc?gct?ggc?atg?tca?act?gca????240

Leu?Lys?Val?Ala?Ile?Ile?Gly?Ala?Gly?Leu?Ala?Gly?Met?Ser?Thr?Ala

65??????????????????70??????????????????75??????????????????80

gtg?gag?ctt?ttg?gat?caa?ggg?cat?gag?gtt?gac?ata?tat?gaa?tcc?aga????288

Val?Glu?Leu?Leu?Asp?Gln?Gly?His?Glu?Val?Asp?Ile?Tyr?Glu?Ser?Arg

85??????????????????90??????????????????95

caa?ttt?att?ggt?ggt?aaa?gtc?ggt?tct?ttt?gta?gat?aag?cgt?gga?aac????336

Gln?Phe?Ile?Gly?Gly?Lys?Val?Gly?Ser?Phe?Val?Asp?Lys?Arg?Gly?Asn

100?????????????????105?????????????????110

cat?att?gaa?atg?gga?ctc?cat?gtg?ttt?ttt?ggt?tgc?tat?aac?aat?ctt????384

His?Ile?Glu?Met?Gly?Leu?His?Val?Phe?Phe?Gly?Cys?Tyr?Asn?Asn?Leu

115?????????????????120?????????????????125

ttc?aga?ctt?atg?aaa?aag?gta?ggt?gca?gat?gaa?aat?tta?ctg?gtg?aag????432

Phe?Arg?Leu?Met?Lys?Lys?Val?Gly?Ala?Asp?Glu?Asn?Leu?Leu?Val?Lys

130?????????????????135?????????????????140

gat?cat?act?cat?acc?ttt?gta?aac?cga?ggt?gga?gaa?att?ggt?gaa?ctt????480

Asp?His?Thr?His?Thr?Phe?Val?Asn?Arg?Gly?Gly?Glu?Ile?Gly?Glu?Leu

145?????????????????150?????????????????155?????????????????160

gat?ttc?cga?ctt?ccg?atg?ggt?gca?cca?tta?cat?ggt?att?cgt?gca?ttt????528

Asp?Phe?Arg?Leu?Pro?Met?Gly?Ala?Pro?Leu?His?Gly?Ile?Arg?Ala?Phe

165?????????????????170?????????????????175

cta?aca?act?aat?caa?ctg?aag?cct?tat?gat?aaa?gca?agg?aat?gct?gtg????576

Leu?Thr?Thr?Asn?Gln?Leu?Lys?Pro?Tyr?Asp?Lys?Ala?Arg?Asn?Ala?Val

180?????????????????185?????????????????190

gct?ctt?gcc?ctt?agc?cca?gtt?gta?cgt?gct?ctt?att?gat?cca?aat?ggt????624

Ala?Leu?Ala?Leu?Ser?Pro?Val?Val?Arg?Ala?Leu?Ile?Asp?Pro?Asn?Gly

195?????????????????200?????????????????205

gca?atg?cag?gat?ata?agg?aac?tta?gat?aat?att?agc?ttt?tct?gat?tgg????672

Ala?Met?Gln?Asp?Ile?Arg?Asn?Leu?Asp?Asn?Ile?Ser?Phe?Ser?Asp?Trp

210?????????????????215?????????????????220

ttc?tta?tcc?aaa?ggc?ggt?acc?cgc?atg?agc?atc?caa?agg?atg?tgg?gat????720

Phe?Leu?Ser?Lys?Gly?Gly?Thr?Arg?Met?Ser?Ile?Gln?Arg?Met?Trp?Asp

225?????????????????230?????????????????235?????????????????240

cca?gtt?gct?tat?gcc?ctc?gga?ttt?att?gac?tgt?gat?aat?atc?agt?gcc????768

Pro?Val?Ala?Tyr?Ala?Leu?Gly?Phe?Ile?Asp?Cys?Asp?Asn?Ile?Ser?Ala

245?????????????????250?????????????????255

cgt?tgt?atg?ctt?act?ata?ttt?tct?cta?ttt?gct?act?aag?aca?gaa?gct????816

Arg?Cys?Met?Leu?Thr?Ile?Phe?Ser?Leu?Phe?Ala?Thr?Lys?Thr?Glu?Ala

260?????????????????265?????????????????270

tct?ctg?ttg?cgt?atg?ttg?aag?ggt?tcg?cct?gat?gtt?tac?tta?agc?ggt????864

Ser?Leu?Leu?Arg?Met?Leu?Lys?Gly?Ser?Pro?Asp?Val?Tyr?Leu?Ser?Gly

275?????????????????280?????????????????285

cct?ata?aga?aag?tat?att?aca?gat?aaa?ggt?gga?agg?ttt?cac?cta?agg????912

Pro?Ile?Arg?Lys?Tyr?Ile?Thr?Asp?Lys?Gly?Gly?Arg?Phe?His?Leu?Arg

290?????????????????295?????????????????300

tgg?ggg?tgt?aga?gag?ata?ctt?tat?gat?gaa?cta?tca?aat?ggc?gac?aca????960

Trp?Gly?Cys?Arg?Glu?Ile?Leu?Tyr?Asp?Glu?Leu?Ser?Asn?Gly?Asp?Thr

305?????????????????310?????????????????315?????????????????320

tat?atc?aca?ggc?att?gca?atg?tcg?aag?gct?acc?aat?aaa?aaa?ctt?gtg????1008

Tyr?Ile?Thr?Gly?Ile?Ala?Met?Ser?Lys?Ala?Thr?Asn?Lys?Lys?Leu?Val

325?????????????????330?????????????????335

aaa?gct?gac?gtg?tat?gtt?gca?gca?tgt?gat?gtt?cct?gga?ata?aaa?agg????1056

Lys?Ala?Asp?Val?Tyr?Val?Ala?Ala?Cys?Asp?Val?Pro?Gly?Ile?Lys?Arg

340?????????????????345?????????????????350

ttg?atc?cca?tcg?gag?tgg?aga?gaa?tgg?gat?cta?ttt?gac?aat?atc?tat????1104

Leu?Ile?Pro?Ser?Glu?Trp?Arg?Glu?Trp?Asp?Leu?Phe?Asp?Asn?Ile?Tyr

355?????????????????360?????????????????365

aaa?cta?gtt?gga?gtt?cca?gtt?gtc?act?gtt?cag?ctt?agg?tac?aat?ggt????1152

Lys?Leu?Val?Gly?Val?Pro?Val?Val?Thr?Val?Gln?Leu?Arg?Tyr?Asn?Gly

370?????????????????375?????????????????380

tgg?gtg?aca?gag?atg?caa?gat?ctg?gaa?aaa?tca?agg?cag?ttg?aga?gct????1200

Trp?Val?Thr?Glu?Met?Gln?Asp?Leu?Glu?Lys?Ser?Arg?Gln?Leu?Arg?Ala

385?????????????????390?????????????????395?????????????????400

gca?gta?gga?ttg?gat?aat?ctt?ctt?tat?act?cca?gat?gca?gac?ttt?tct????1248

Ala?Val?Gly?Leu?Asp?Asn?Leu?Leu?Tyr?Thr?Pro?Asp?Ala?Asp?Phe?Ser

405?????????????????410?????????????????415

tgt?ttt?tct?gat?ctt?gca?ctc?tcg?tcg?cct?gaa?gat?tat?tat?att?gaa????1296

Cys?Phe?Ser?Asp?Leu?Ala?Leu?Ser?Ser?Pro?Glu?Asp?Tyr?Tyr?Ile?Glu

420?????????????????425?????????????????430

gga?caa?ggg?tcc?cta?ata?cag?gct?gtt?ctc?acg?cca?ggg?gat?cca?tac????1344

Gly?Gln?Gly?Ser?Leu?Ile?Gln?Ala?Val?Leu?Thr?Pro?Gly?Asp?Pro?Tyr

435?????????????????440?????????????????445

atg?ccc?cta?cct?aat?gat?gca?att?ata?gaa?aga?gtt?cgg?aaa?cag?gtt????1392

Met?Pro?Leu?Pro?Asn?Asp?Ala?Ile?Ile?Glu?Arg?Val?Arg?Lys?Gln?Val

450?????????????????455?????????????????460

ttg?gat?tta?ttc?cca?tcc?tct?caa?ggc?ctg?gaa?gtt?cta?tgg?tct?tcg????1440

Leu?Asp?Leu?Phe?Pro?Ser?Ser?Gln?Gly?Leu?Glu?Val?Leu?Trp?Ser?Ser

465?????????????????470?????????????????475?????????????????480

gtg?gtt?aaa?atc?gga?caa?tcc?cta?tat?cgg?gag?ggg?cct?gga?aag?gac????1488

Val?Val?Lys?Ile?Gly?Gln?Ser?Leu?Tyr?Arg?Glu?Gly?Pro?Gly?Lys?Asp

485?????????????????490?????????????????495

cca?ttc?aga?cct?gat?cag?aag?aca?cca?gta?aaa?aat?ttc?ttc?ctt?gca????1536

Pro?Phe?Arg?Pro?Asp?Gln?Lys?Thr?Pro?Val?Lys?Asn?Phe?Phe?Leu?Ala

500?????????????????505?????????????????510

ggt?tca?tac?acc?aaa?cag?gat?tac?att?gac?agt?atg?gaa?gga?gcg?acc????1584

Gly?Ser?Tyr?Thr?Lys?Gln?Asp?Tyr?Ile?Asp?Ser?Met?Glu?Gly?Ala?Thr

515?????????????????520?????????????????525

cta?tcg?ggg?aga?caa?gca?gct?gca?tat?atc?tgc?agc?gcc?ggt?gaa?gat????1632

Leu?Ser?Gly?Arg?Gln?Ala?Ala?Ala?Tyr?Ile?Cys?Ser?Ala?Gly?Glu?Asp

530?????????????????535?????????????????540

ctg?gca?gca?ctt?cgc?aag?aag?atc?gct?gct?gat?cat?cca?gag?caa?ctg????1680

Leu?Ala?Ala?Leu?Arg?Lys?Lys?Ile?Ala?Ala?Asp?His?Pro?Glu?Gln?Leu

545?????????????????550?????????????????555?????????????????560

atc?aac?aaa?gat?tct?aac?gtg?tcg?gat?gaa?ctg?agt?ctc?gta?taa????????1725

Ile?Asn?Lys?Asp?Ser?Asn?Val?Ser?Asp?Glu?Leu?Ser?Leu?Val

565?????????????????570

<210>132

<211>574

<212>PRT

＜213〉daffodil

<400>132

Met?Ala?Ser?Ser?Thr?Cys?Leu?Ile?His?Ser?Ser?Ser?Phe?Gly?Val?Gly

1???????????????5???????????????????10??????????????????15

Gly?Lys?Lys?Val?Lys?Met?Asn?Thr?Met?Ile?Arg?Ser?Lys?Leu?Phe?Ser

20??????????????????25??????????????????30

Ile?Arg?Ser?Ala?Leu?Asp?Thr?Lys?Val?Ser?Asp?Met?Ser?Val?Asn?Ala

35??????????????????40??????????????????45

Pro?Lys?Gly?Leu?Phe?Pro?Pro?Glu?Pro?Glu?His?Tyr?Arg?Gly?Pro?Lys

50??????????????????55??????????????????60

Leu?Lys?Val?Ala?Ile?Ile?Gly?Ala?Gly?Leu?Ala?Gly?Met?Ser?Thr?Ala

65??????????????????70??????????????????75??????????????????80

Val?Glu?Leu?Leu?Asp?Gln?Gly?His?Glu?Val?Asp?Ile?Tyr?Glu?Ser?Arg

85??????????????????90??????????????????95

Gln?Phe?Ile?Gly?Gly?Lys?Val?Gly?Ser?Phe?Val?Asp?Lys?Arg?Gly?Asn

100?????????????????105?????????????????110

His?Ile?Glu?Met?Gly?Leu?His?Val?Phe?Phe?Gly?Cys?Tyr?Asn?Asn?Leu

115?????????????????120?????????????????125

Phe?Arg?Leu?Met?Lys?Lys?Val?Gly?Ala?Asp?Glu?Asn?Leu?Leu?Val?Lys

130?????????????????135?????????????????140

Asp?His?Thr?His?Thr?Phe?Val?Asn?Arg?Gly?Gly?Glu?Ile?Gly?Glu?Leu

145?????????????????150?????????????????155?????????????????160

Asp?Phe?Arg?Leu?Pro?Met?Gly?Ala?Pro?Leu?His?Gly?Ile?Arg?Ala?Phe

165?????????????????170?????????????????175

Leu?Thr?Thr?Asn?Gln?Leu?Lys?Pro?Tyr?Asp?Lys?Ala?Arg?Asn?Ala?Val

180?????????????????185?????????????????190

Ala?Leu?Ala?Leu?Ser?Pro?Val?Val?Arg?Ala?Leu?Ile?Asp?Pro?Asn?Gly

195?????????????????200?????????????????205

Ala?Met?Gln?Asp?Ile?Arg?Asn?Leu?Asp?Asn?Ile?Ser?Phe?Ser?Asp?Trp

210?????????????????215?????????????????220

Phe?Leu?Ser?Lys?Gly?Gly?Thr?Arg?Met?Ser?Ile?Gln?Arg?Met?Trp?Asp

225?????????????????230?????????????????235?????????????????240

Pro?Val?Ala?Tyr?Ala?Leu?Gly?Phe?Ile?Asp?Cys?Asp?Asn?Ile?Ser?Ala

245?????????????????250?????????????????255

Arg?Cys?Met?Leu?Thr?Ile?Phe?Ser?Leu?Phe?Ala?Thr?Lys?Thr?Glu?Ala

260?????????????????265?????????????????270

Ser?Leu?Leu?Arg?Met?Leu?Lys?Gly?Ser?Pro?Asp?Val?Tyr?Leu?Ser?Gly

275?????????????????280?????????????????285

Pro?Ile?Arg?Lys?Tyr?Ile?Thr?Asp?Lys?Gly?Gly?Arg?Phe?His?Leu?Arg

290?????????????????295?????????????????300

Trp?Gly?Cys?Arg?Glu?Ile?Leu?Tyr?Asp?Glu?Leu?Ser?Asn?Gly?Asp?Thr

305?????????????????310?????????????????315?????????????????320

Tyr?Ile?Thr?Gly?Ile?Ala?Met?Ser?Lys?Ala?Thr?Asn?Lys?Lys?Leu?Val

325?????????????????330?????????????????335

Lys?Ala?Asp?Val?Tyr?Val?Ala?Ala?Cys?Asp?Val?Pro?Gly?Ile?Lys?Arg

340?????????????????345?????????????????350

Leu?Ile?Pro?Ser?Glu?Trp?Arg?Glu?Trp?Asp?Leu?Phe?Asp?Asn?Ile?Tyr

355?????????????????360?????????????????365

Lys?Leu?Val?Gly?Val?Pro?Val?Val?Thr?Val?Gln?Leu?Arg?Tyr?Asn?Gly

370?????????????????375?????????????????380

Trp?Val?Thr?Glu?Met?Gln?Asp?Leu?Glu?Lys?Ser?Arg?Gln?Leu?Arg?Ala

385?????????????????390?????????????????395?????????????????400

Ala?Val?Gly?Leu?Asp?Asn?Leu?Leu?Tyr?Thr?Pro?Asp?Ala?Asp?Phe?Ser

405?????????????????410?????????????????415

Cys?Phe?Ser?Asp?Leu?Ala?Leu?Ser?Ser?Pro?Glu?Asp?Tyr?Tyr?Ile?Glu

420?????????????????425?????????????????430

Gly?Gln?Gly?Ser?Leu?Ile?Gln?Ala?Val?Leu?Thr?Pro?Gly?Asp?Pro?Tyr

435?????????????????440?????????????????445

Met?Pro?Leu?Pro?Asn?Asp?Ala?Ile?Ile?Glu?Arg?Val?Arg?Lys?Gln?Val

450?????????????????455?????????????????460

Leu?Asp?Leu?Phe?Pro?Ser?Ser?Gln?Gly?Leu?Glu?Val?Leu?Trp?Ser?Ser

465?????????????????470?????????????????475?????????????????480

Val?Val?Lys?Ile?Gly?Gln?Ser?Leu?Tyr?Arg?Glu?Gly?Pro?Gly?Lys?Asp

485?????????????????490?????????????????495

Pro?Phe?Arg?Pro?Asp?Gln?Lys?Thr?Pro?Val?Lys?Asn?Phe?Phe?Leu?Ala

500?????????????????505?????????????????510

Gly?Ser?Tyr?Thr?Lys?Gln?Asp?Tyr?Ile?Asp?Ser?Met?Glu?Gly?Ala?Thr

515?????????????????520?????????????????525

Leu?Ser?Gly?Arg?Gln?Ala?Ala?Ala?Tyr?Ile?Cys?Ser?Ala?Gly?Glu?Asp

530?????????????????535?????????????????540

Leu?Ala?Ala?Leu?Arg?Lys?Lys?Ile?Ala?Ala?Asp?His?Pro?Glu?Gln?Leu

545?????????????????550?????????????????555?????????????????560

Ile?Asn?Lys?Asp?Ser?Asn?Val?Ser?Asp?Glu?Leu?Ser?Leu?Val

565?????????????????570

<210>133

<211>1848

<212>DNA

＜213〉edible tomato

<220>

<221>CDS

<222>(1)..(1848)

<223>

<400>133

atg?tgt?acc?ttg?agt?ttt?atg?tat?cct?aat?tca?ctt?ctt?gat?ggt?acc????48

Met?Cys?Thr?Leu?Ser?Phe?Met?Tyr?Pro?Asn?Ser?Leu?Leu?Asp?Gly?Thr

1???????????????5???????????????????10??????????????????15

tgc?aag?act?gta?gct?ttg?ggt?gat?agc?aaa?cca?aga?tac?aat?aaa?cag????96

Cys?Lys?Thr?Val?Ala?Leu?Gly?Asp?Ser?Lys?Pro?Arg?Tyr?Asn?Lys?Gln

20??????????????????25??????????????????30

aga?agt?tct?tgt?ttt?gac?cct?ttg?ata?att?gga?aat?tgt?act?gat?cag????144

Arg?Ser?Ser?Cys?Phe?Asp?Pro?Leu?Ile?Ile?Gly?Asn?Cys?Thr?Asp?Gln

35??????????????????40??????????????????45

cag?cag?ctt?tgt?ggc?ttg?agt?tgg?ggg?gtg?gac?aag?gct?aag?gga?aga????192

Gln?Gln?Leu?Cys?Gly?Leu?Ser?Trp?Gly?Val?Asp?Lys?Ala?Lys?Gly?Arg

50??????????????????55??????????????????60

aga?ggg?ggt?act?gtt?tcc?aat?ttg?aaa?gca?gtt?gta?gat?gta?gac?aaa????240

Arg?Gly?Gly?Thr?Val?Ser?Asn?Leu?Lys?Ala?Val?Val?Asp?Val?Asp?Lys

65??????????????????70??????????????????75??????????????????80

aga?gtg?gag?agc?tat?ggc?agt?agt?gat?gta?gaa?gga?aat?gag?agt?ggc????288

Arg?Val?Glu?Ser?Tyr?Gly?Ser?Ser?Asp?Val?Glu?Gly?Asn?Glu?Ser?Gly

85??????????????????90??????????????????95

agc?tat?gat?gcc?att?gtt?ata?ggt?tca?gga?ata?ggt?gga?ttg?gtg?gca????336

Ser?Tyr?Asp?Ala?Ile?Val?Ile?Gly?Ser?Gly?Ile?Gly?Gly?Leu?Val?Ala

100?????????????????105?????????????????110

gcg?acg?cag?ctg?gcg?gtt?aag?gga?gct?aag?gtt?tta?gtt?ctg?gag?aag????384

Ala?Thr?Gln?Leu?Ala?Val?Lys?Gly?Ala?Lys?Val?Leu?Val?Leu?Glu?Lys

115?????????????????120?????????????????125

tat?gtt?att?cct?ggt?gga?agc?tct?ggc?ttt?tac?gag?agg?gat?ggt?tat????432

Tyr?Val?Ile?Pro?Gly?Gly?Ser?Ser?Gly?Phe?Tyr?Glu?Arg?Asp?Gly?Tyr

130?????????????????135?????????????????140

aag?ttt?gat?gtt?ggt?tca?tca?gtg?atg?ttt?gga?ttc?agt?gat?aag?gga????480

Lys?Phe?Asp?Val?Gly?Ser?Ser?Val?Met?Phe?Gly?Phe?Ser?Asp?Lys?Gly

145?????????????????150?????????????????155?????????????????160

aac?ctc?aat?tta?att?act?caa?gca?ttg?gca?gca?gta?gga?cgt?aaa?tta????528

Asn?Leu?Asn?Leu?Ile?Thr?Gln?Ala?Leu?Ala?Ala?Val?Gly?Arg?Lys?Leu

165?????????????????170?????????????????175

gaa?gtt?ata?cct?gac?cca?aca?act?gta?cat?ttc?cac?ctg?cca?aat?gac????576

Glu?Val?Ile?Pro?Asp?Pro?Thr?Thr?Val?His?Phe?His?Leu?Pro?Asn?Asp

180?????????????????185?????????????????190

ctt?tct?gtt?cgt?ata?cac?cga?gag?tat?gat?gac?ttc?att?gaa?gag?ctt????624

Leu?Ser?Val?Arg?Ile?His?Arg?Glu?Tyr?Asp?Asp?Phe?Ile?Glu?Glu?Leu

195?????????????????200?????????????????205

gtg?agt?aaa?ttt?cca?cat?gaa?aag?gaa?ggg?att?atc?aaa?ttt?tac?agt????672

Val?Ser?Lys?Phe?Pro?His?Glu?Lys?Glu?Gly?Ile?Ile?Lys?Phe?Tyr?Ser

210?????????????????215?????????????????220

gaa?tgc?tgg?aag?atc?ttt?aat?tct?ctg?aat?tca?ttg?gaa?ctg?aag?tct????720

Glu?Cys?Trp?Lys?Ile?Phe?Asn?Ser?Leu?Asn?Ser?Leu?Glu?Leu?Lys?Ser

225?????????????????230?????????????????235?????????????????240

ttg?gag?gaa?ccc?atc?tac?ctt?ttt?ggc?cag?ttc?ttt?aag?aag?ccc?ctt?????768

Leu?Glu?Glu?Pro?Ile?Tyr?Leu?Phe?Gly?Gln?Phe?Phe?Lys?Lys?Pro?Leu

245?????????????????250?????????????????255

gaa?tgc?ttg?act?ctt?gcc?tac?tat?ttg?ccc?cag?aat?gct?ggt?agc?atc?????816

Glu?Cys?Leu?Thr?Leu?Ala?Tyr?Tyr?Leu?Pro?Gln?Asn?Ala?Gly?Ser?Ile

260?????????????????265?????????????????270

gct?cgg?aag?tat?ata?aga?gat?cct?ggg?ttg?ctg?tct?ttt?ata?gat?gca?????864

Ala?Arg?Lys?Tyr?Ile?Arg?Asp?Pro?Gly?Leu?Leu?Ser?Phe?Ile?Asp?Ala

275?????????????????280?????????????????285

gag?tgc?ttt?atc?gtg?agt?aca?gtt?aat?gca?tta?caa?aca?cca?atg?atc?????912

Glu?Cys?Phe?Ile?Val?Ser?Thr?Val?Asn?Ala?Leu?Gln?Thr?Pro?Met?Ile

290?????????????????295?????????????????300

aat?gca?agc?atg?gtt?cta?tgt?gac?aga?cat?ttt?ggc?gga?atc?aac?tac?????960

Asn?Ala?Ser?Met?Val?Leu?Cys?Asp?Arg?His?Phe?Gly?Gly?Ile?Asn?Tyr

305?????????????????310?????????????????315?????????????????320

ccc?gtt?ggt?gga?gtt?ggc?gag?atc?gcc?aaa?tcc?tta?gca?aaa?ggc?ttg????1008

Pro?Val?Gly?Gly?Val?Gly?Glu?Ile?Ala?Lys?Ser?Leu?Ala?Lys?Gly?Leu

325?????????????????330?????????????????335

gat?gat?cac?gga?agt?cag?ata?ctt?tat?agg?gca?aat?gtt?aca?agt?atc????1056

Asp?Asp?His?Gly?Ser?Gln?Ile?Leu?Tyr?Arg?Ala?Asn?Val?Thr?Ser?Ile

340?????????????????345?????????????????350

att?ttg?gac?aat?ggc?aaa?gct?gtg?gga?gtg?aag?ctt?tct?gac?ggg?agg????1104

Ile?Leu?Asp?Asn?Gly?Lys?Ala?Val?Gly?Val?Lys?Leu?Ser?Asp?Gly?Arg

355?????????????????360?????????????????365

aag?ttt?tat?gct?aaa?acc?ata?gta?tcg?aat?gct?acc?aga?tgg?gat?act????1152

Lys?Phe?Tyr?Ala?Lys?Thr?Ile?Val?Ser?Asn?Ala?Thr?Arg?Trp?Asp?Thr

370?????????????????375?????????????????380

ttt?gga?aag?ctt?tta?aaa?gct?gag?aat?ctg?cca?aaa?gaa?gaa?gaa?aat????1200

Phe?Gly?Lys?Leu?Leu?Lys?Ala?Glu?Asn?Leu?Pro?Lys?Glu?Glu?Glu?Asn

385?????????????????390?????????????????395?????????????????400

ttc?cag?aaa?gct?tat?gta?aaa?gca?cct?tct?ttt?ctt?tct?att?cat?atg????1248

Phe?Gln?Lys?Ala?Tyr?Val?Lys?Ala?Pro?Ser?Phe?Leu?Ser?Ile?His?Met

405?????????????????410?????????????????415

gga?gtt?aaa?gca?gat?gta?ctc?cca?cca?gac?aca?gat?tgt?cac?cat?ttt????1296

Gly?Val?Lys?Ala?Asp?Val?Leu?Pro?Pro?Asp?Thr?Asp?Cys?His?His?Phe

420?????????????????425?????????????????430

gtc?ctc?gag?gat?gat?tgg?aca?aat?ttg?gag?aaa?cca?tat?gga?agt?ata????1344

Val?Leu?Glu?Asp?Asp?Trp?Thr?Asn?Leu?Glu?Lys?Pro?Tyr?Gly?Ser?Ile

435?????????????????440?????????????????445

ttc?ttg?agt?att?cca?aca?gtt?ctt?gat?tcc?tca?ttg?gcc?cca?gaa?gga????1392

Phe?Leu?Ser?Ile?Pro?Thr?Val?Leu?Asp?Ser?Ser?Leu?Ala?Pro?Glu?Gly

450?????????????????455?????????????????460

cac?cat?att?ctt?cac?att?ttt?aca?aca?tcg?agc?att?gaa?gat?tgg?gag????1440

His?His?Ile?Leu?His?Ile?Phe?Thr?Thr?Ser?Ser?Ile?Glu?Asp?Trp?Glu

465?????????????????470?????????????????475?????????????????480

gga?ctc?tct?ccg?aaa?gac?tat?gaa?gcg?aag?aaa?gag?gtt?gtt?gct?gaa????1488

Gly?Leu?Ser?Pro?Lys?Asp?Tyr?Glu?Ala?Lys?Lys?Glu?Val?Val?Ala?Glu

485?????????????????490?????????????????495

agg?att?ata?agc?aga?ctt?gaa?aaa?aca?ctc?ttc?cca?ggg?ctt?aag?tca????1536

Arg?Ile?Ile?Ser?Arg?Leu?Glu?Lys?Thr?Leu?Phe?Pro?Gly?Leu?Lys?Ser

500?????????????????505?????????????????510

tct?att?ctc?ttt?aag?gag?gtg?gga?act?cca?aag?acc?cac?aga?cga?tac????1584

Ser?Ile?Leu?Phe?Lys?Glu?Val?Gly?Thr?Pro?Lys?Thr?His?Arg?Arg?Tyr

515?????????????????520?????????????????525

ctt?gct?cgt?gat?agt?ggt?acc?tat?gga?cca?atg?cca?cgc?gga?aca?cct????1632

Leu?Ala?Arg?Asp?Ser?Gly?Thr?Tyr?Gly?Pro?Met?Pro?Arg?Gly?Thr?Pro

530?????????????????535?????????????????540

aag?gga?ctc?ctg?gga?atg?cct?ttc?aat?acc?act?gct?ata?gat?ggt?cta????1680

Lys?Gly?Leu?Leu?Gly?Met?Pro?Phe?Asn?Thr?Thr?Ala?Ile?Asp?Gly?Leu

545?????????????????550?????????????????555?????????????????560

tat?tgt?gtt?ggc?gat?agt?tgc?ttc?cca?gga?caa?ggt?gtt?ata?gct?gta????1728

Tyr?Cys?Val?Gly?Asp?Ser?Cys?Phe?Pro?Gly?Gln?Gly?Val?Ile?Ala?Val

565?????????????????570?????????????????575

gcc?ttt?tca?gga?gta?atg?tgc?gct?cat?cgt?gtt?gca?gct?gac?tta?ggg????1776

Ala?Phe?Ser?Gly?Val?Met?Cys?Ala?His?Arg?Val?Ala?Ala?Asp?Leu?Gly

580?????????????????585?????????????????590

ttt?gaa?aaa?aaa?tca?gat?gtg?ctg?gac?agt?gct?ctt?ctt?aga?cta?ctt????1824

Phe?Glu?Lys?Lys?Ser?Asp?Val?Leu?Asp?Ser?Ala?Leu?Leu?Arg?Leu?Leu

595?????????????????600?????????????????605

ggt?tgg?tta?agg?aca?cta?gca?tga????????????????????????????????????1848

Gly?Trp?Leu?Arg?Thr?Leu?Ala

610?????????????????615

<210>134

<211>615

<212>PRT

＜213〉edible tomato

<400>134

Met?Cys?Thr?Leu?Ser?Phe?Met?Tyr?Pro?Asn?Ser?Leu?Leu?Asp?Gly?Thr

1???????????????5???????????????????10??????????????????15

Cys?Lys?Thr?Val?Ala?Leu?Gly?Asp?Ser?Lys?Pro?Arg?Tyr?Asn?Lys?Gln

20??????????????????25??????????????????30

Arg?Ser?Ser?Cys?Phe?Asp?Pro?Leu?Ile?Ile?Gly?Asn?Cys?Thr?Asp?Gln

35??????????????????40??????????????????45

Gln?Gln?Leu?Cys?Gly?Leu?Ser?Trp?Gly?Val?Asp?Lys?Ala?Lys?Gly?Arg

50??????????????????55??????????????????60

Arg?Gly?Gly?Thr?Val?Ser?Asn?Leu?Lys?Ala?Val?Val?Asp?Val?Asp?Lys

65??????????????????70??????????????????75??????????????????80

Arg?Val?Glu?Ser?Tyr?Gly?Ser?Ser?Asp?Val?Glu?Gly?Asn?Glu?Ser?Gly

85??????????????????90??????????????????95

Ser?Tyr?Asp?Ala?Ile?Val?Ile?Gly?Ser?Gly?Ile?Gly?Gly?Leu?Val?Ala

100?????????????????105?????????????????110

Ala?Thr?Gln?Leu?Ala?Val?Lys?Gly?Ala?Lys?Val?Leu?Val?Leu?Glu?Lys

115?????????????????120?????????????????125

Tyr?Val?Ile?Pro?Gly?Gly?Ser?Ser?Gly?Phe?Tyr?Glu?Arg?Asp?Gly?Tyr

130?????????????????135?????????????????140

Lys?Phe?Asp?Val?Gly?Ser?Ser?Val?Met?Phe?Gly?Phe?Ser?Asp?Lys?Gly

145?????????????????150?????????????????155?????????????????160

Asn?Leu?Asn?Leu?Ile?Thr?Gln?Ala?Leu?Ala?Ala?Val?Gly?Arg?Lys?Leu

165?????????????????170?????????????????175

Glu?Val?Ile?Pro?Asp?Pro?Thr?Thr?Val?His?Phe?His?Leu?Pro?Asn?Asp

180?????????????????185?????????????????190

Leu?Ser?Val?Arg?Ile?His?Arg?Glu?Tyr?Asp?Asp?Phe?Ile?Glu?Glu?Leu

195?????????????????200?????????????????205

Val?Ser?Lys?Phe?Pro?His?Glu?Lys?Glu?Gly?Ile?Ile?Lys?Phe?Tyr?Ser

210?????????????????215?????????????????220

Glu?Cys?Trp?Lys?Ile?Phe?Asn?Ser?Leu?Asn?Ser?Leu?Glu?Leu?Lys?Ser

225?????????????????230?????????????????235?????????????????240

Leu?Glu?Glu?Pro?Ile?Tyr?Leu?Phe?Gly?Gln?Phe?Phe?Lys?Lys?Pro?Leu

245?????????????????250?????????????????255

Glu?Cys?Leu?Thr?Leu?Ala?Tyr?Tyr?Leu?Pro?Gln?Asn?Ala?Gly?Ser?Ile

260?????????????????265?????????????????270

Ala?Arg?Lys?Tyr?Ile?Arg?Asp?Pro?Gly?Leu?Leu?Ser?Phe?Ile?Asp?Ala

275?????????????????280?????????????????285

Glu?Cys?Phe?Ile?Val?Ser?Thr?Val?Asn?Ala?Leu?Gln?Thr?Pro?Met?Ile

290?????????????????295?????????????????300

Asn?Ala?Ser?Met?Val?Leu?Cys?Asp?Arg?His?Phe?Gly?Gly?Ile?Asn?Tyr

305?????????????????310?????????????????315?????????????????320

Pro?Val?Gly?Gly?Val?Gly?Glu?Ile?Ala?Lys?Ser?Leu?Ala?Lys?Gly?Leu

325?????????????????330?????????????????335

Asp?Asp?His?Gly?Ser?Gln?Ile?Leu?Tyr?Arg?Ala?Asn?Val?Thr?Ser?Ile

340?????????????????345?????????????????350

Ile?Leu?Asp?Asn?Gly?Lys?Ala?Val?Gly?Val?Lys?Leu?Ser?Asp?Gly?Arg

355?????????????????360?????????????????365

Lys?Phe?Tyr?Ala?Lys?Thr?Ile?Val?Ser?Asn?Ala?Thr?Arg?Trp?Asp?Thr

370?????????????????375?????????????????380

Phe?Gly?Lys?Leu?Leu?Lys?Ala?Glu?Asn?Leu?Pro?Lys?Glu?Glu?Glu?Asn

385?????????????????390?????????????????395?????????????????400

Phe?Gln?Lys?Ala?Tyr?Val?Lys?Ala?Pro?Ser?Phe?Leu?Ser?Ile?His?Met

405?????????????????410?????????????????415

Gly?Val?Lys?Ala?Asp?Val?Leu?Pro?Pro?Asp?Thr?Asp?Cys?His?His?Phe

420?????????????????425?????????????????430

Val?Leu?Glu?Asp?Asp?Trp?Thr?Asn?Leu?Glu?Lys?Pro?Tyr?Gly?Ser?Ile

435?????????????????440?????????????????445

Phe?Leu?Ser?Ile?Pro?Thr?Val?Leu?Asp?Ser?Ser?Leu?Ala?Pro?Glu?Gly

450?????????????????455?????????????????460

His?His?Ile?Leu?His?Ile?Phe?Thr?Thr?Ser?Ser?Ile?Glu?Asp?Trp?Glu

465?????????????????470?????????????????475?????????????????480

Gly?Leu?Ser?Pro?Lys?Asp?Tyr?Glu?Ala?Lys?Lys?Glu?Val?Val?Ala?Glu

485?????????????????490?????????????????495

Arg?Ile?Ile?Ser?Arg?Leu?Glu?Lys?Thr?Leu?Phe?Pro?Gly?Leu?Lys?Ser

500?????????????????505?????????????????510

Ser?Ile?Leu?Phe?Lys?Glu?Val?Gly?Thr?Pro?Lys?Thr?His?Arg?Arg?Tyr

515?????????????????520?????????????????525

Leu?Ala?Arg?Asp?Ser?Gly?Thr?Tyr?Gly?Pro?Met?Pro?Arg?Gly?Thr?Pro

530?????????????????535?????????????????540

Lys?Gly?Leu?Leu?Gly?Met?Pro?Phe?Asn?Thr?Thr?Ala?Ile?Asp?Gly?Leu

545?????????????????550?????????????????555?????????????????560

Tyr?Cys?Val?Gly?Asp?Ser?Cys?Phe?Pro?Gly?Gln?Gly?Val?Ile?Ala?Val

565?????????????????570?????????????????575

Ala?Phe?Ser?Gly?Val?Met?Cys?Ala?His?Arg?Val?Ala?Ala?Asp?Leu?Gly

580?????????????????585?????????????????590

Phe?Glu?Lys?Lys?Ser?Asp?Val?Leu?Asp?Ser?Ala?Leu?Leu?Arg?Leu?Leu

595?????????????????600?????????????????605

Gly?Trp?Leu?Arg?Thr?Leu?Ala

610?????????????????615

<210>135

<211>1233

<212>DNA

＜213〉marigold

<220>

<221>CDS

<222>(1)..(1233)

<223>

<400>135

atg?gcc?aca?cac?aaa?ctc?ctt?caa?ttc?acc?acc?aat?ctc?cca?cca?tct????48

Met?Ala?Thr?His?Lys?Leu?Leu?Gln?Phe?Thr?Thr?Asn?Leu?Pro?Pro?Ser

1???????????????5???????????????????10??????????????????15

tct?tct?tca?atc?tct?act?ggc?tgt?tca?ctc?tcc?ccc?ttc?ttc?ctc?aaa????96

Ser?Ser?Ser?Ile?Ser?Thr?Gly?Cys?Ser?Leu?Ser?Pro?Phe?Phe?Leu?Lys

20??????????????????25??????????????????30

tca?tct?tct?cat?tcc?cct?aac?cct?cgc?cga?cac?cgc?cgc?tcc?gcc?gta????144

Ser?Ser?Ser?His?Ser?Pro?Asn?Pro?Arg?Arg?His?Arg?Arg?Ser?Ala?Val

35??????????????????40??????????????????45

tgc?tgc?tct?ttc?gcc?tca?ctc?gac?tct?gca?aaa?atc?aaa?gtc?gtt?ggc????192

Cys?Cys?Ser?Phe?Ala?Ser?Leu?Asp?Ser?Ala?Lys?Ile?Lys?Val?Val?Gly

50??????????????????55??????????????????60

gtc?ggt?ggt?ggt?ggc?aac?aat?gcc?gtt?aac?cgc?atg?att?ggt?agc?ggc????240

Val?Gly?Gly?Gly?Gly?Asn?Asn?Ala?Val?Asn?Arg?Met?Ile?Gly?Ser?Gly

65??????????????????70??????????????????75??????????????????80

tta?cag?ggt?gtt?gat?ttt?tac?gcc?att?aac?acg?gac?tca?caa?gcg?ctt????288

Leu?Gln?Gly?Val?Asp?Phe?Tyr?Ala?Ile?Asn?Thr?Asp?Ser?Gln?Ala?Leu

85??????????????????90??????????????????95

ctg?caa?tct?gtt?gca?cat?aac?cct?att?caa?att?ggg?gag?ctt?ttg?act????336

Leu?Gln?Ser?Val?Ala?His?Asn?Pro?Ile?Gln?Ile?Gly?Glu?Leu?Leu?Thr

100?????????????????105?????????????????110

cgt?gga?tta?ggt?act?ggt?ggg?aac?ccg?ctt?ttg?gga?gaa?cag?gct?gcg????384

Arg?Gly?Leu?Gly?Thr?Gly?Gly?Asn?Pro?Leu?Leu?Gly?Glu?Gln?Ala?Ala

115?????????????????120?????????????????125

gag?gag?tcg?aag?gaa?gcg?att?ggg?aat?gcg?ctt?aaa?ggg?tcg?gat?ctt????432

Glu?Glu?Ser?Lys?Glu?Ala?Ile?Gly?Asn?Ala?Leu?Lys?Gly?Ser?Asp?Leu

130?????????????????135?????????????????140

gtg?ttt?ata?aca?gca?ggt?atg?ggt?ggt?ggg?acg?ggt?tcg?ggt?gct?gct????480

Val?Phe?Ile?Thr?Ala?Gly?Met?Gly?Gly?Gly?Thr?Gly?Ser?Gly?Ala?Ala

145?????????????????150?????????????????155?????????????????160

cca?gtt?gta?gcg?cag?ata?gcg?aaa?gaa?gca?ggg?tat?tta?act?gtt?ggt????528

Pro?Val?Val?Ala?Gln?Ile?Ala?Lys?Glu?Ala?Gly?Tyr?Leu?Thr?Val?Gly

165?????????????????170?????????????????175

gtt?gta?acg?tac?cca?ttc?agc?ttt?gaa?ggc?cgt?aaa?aga?tca?gta?cag????576

Val?Val?Thr?Tyr?Pro?Phe?Ser?Phe?Glu?Gly?Arg?Lys?Arg?Ser?Val?Gln

180?????????????????185?????????????????190

gcg?tta?gag?gct?att?gag?aag?ctg?caa?aag?aac?gtt?gac?aca?ctt?ata?????624

Ala?Leu?Glu?Ala?Ile?Glu?Lys?Leu?Gln?Lys?Asn?Val?Asp?Thr?Leu?Ile

195?????????????????200?????????????????205

gtg?att?cca?aat?gac?cgt?ttg?ctg?gat?att?gct?gat?gaa?aac?acg?cct?????672

Val?Ile?Pro?Asn?Asp?Arg?Leu?Leu?Asp?Ile?Ala?Asp?Glu?Asn?Thr?Pro

210?????????????????215?????????????????220

ctt?cag?gat?gct?ttt?ctt?ctt?gct?gat?gat?gta?ctc?cgc?caa?gga?gtt?????720

Leu?Gln?Asp?Ala?Phe?Leu?Leu?Ala?Asp?Asp?Val?Leu?Arg?Gln?Gly?Val

225?????????????????230?????????????????235?????????????????240

caa?gga?atc?tca?gat?ata?att?aca?ata?cct?ggg?ctg?gta?aat?gtg?gac?????768

Gln?Gly?Ile?Ser?Asp?Ile?Ile?Thr?Ile?Pro?Gly?Leu?Val?Asn?Val?Asp

245?????????????????250?????????????????255

ttt?gca?gac?gtt?aaa?gca?gtc?atg?aaa?gat?tct?gga?act?gca?atg?ctt?????816

Phe?Ala?Asp?Val?Lys?Ala?Val?Met?Lys?Asp?Ser?Gly?Thr?Ala?Met?Leu

260?????????????????265?????????????????270

ggt?gtc?ggt?gtt?tcc?tca?agt?aaa?aac?cga?gct?gaa?gaa?gca?gct?gaa?????864

Gly?Val?Gly?Val?Ser?Ser?Ser?Lys?Asn?Arg?Ala?Glu?Glu?Ala?Ala?Glu

275?????????????????280?????????????????285

caa?gca?act?ctt?gct?cct?ttg?att?gga?tca?tca?att?caa?tct?gct?aca?????912

Gln?Ala?Thr?Leu?Ala?Pro?Leu?Ile?Gly?Ser?Ser?Ile?Gln?Ser?Ala?Thr

290?????????????????295?????????????????300

ggt?gtt?gtt?tat?aat?att?acc?gga?ggg?aag?gac?ata?act?cta?caa?gaa?????960

Gly?Val?Val?Tyr?Asn?Ile?Thr?Gly?Gly?Lys?Asp?Ile?Thr?Leu?Gln?Glu

305?????????????????310?????????????????315?????????????????320

gtc?aac?agg?gtt?tct?cag?gtg?gta?aca?agt?ttg?gca?gat?cca?tca?gca????1008

Val?Asn?Arg?Val?Ser?Gln?Val?Val?Thr?Ser?Leu?Ala?Asp?Pro?Ser?Ala

325?????????????????330?????????????????335

aac?att?ata?ttc?ggg?gca?gtg?gta?gat?gag?aga?tac?aac?ggg?gag?att????1056

Asn?Ile?Ile?Phe?Gly?Ala?Val?Val?Asp?Glu?Arg?Tyr?Asn?Gly?Glu?Ile

340?????????????????345?????????????????350

cat?gtg?acc?att?gtt?gct?act?ggc?ttt?gcc?cag?tcg?ttt?cag?aaa?tct????1104

His?Val?Thr?Ile?Val?Ala?Thr?Gly?Phe?Ala?Gln?Ser?Phe?Gln?Lys?Ser

355?????????????????360?????????????????365

ctt?ctt?gct?gac?ccg?aaa?gga?gca?aaa?ctt?gtt?gat?aga?aat?caa?gaa????1152

Leu?Leu?Ala?Asp?Pro?Lys?Gly?Ala?Lys?Leu?Val?Asp?Arg?Asn?Gln?Glu

370?????????????????375?????????????????380

cct?aca?caa?cct?ttg?act?tcc?gcg?aga?tct?ttg?aca?aca?cct?tct?cct????1200

Pro?Thr?Gln?Pro?Leu?Thr?Ser?Ala?Arg?Ser?Leu?Thr?Thr?Pro?Ser?Pro

385?????????????????390?????????????????395?????????????????400

gct?ccg?tct?cgg?tct?agg?aaa?ctc?ttc?ttt?taa????????????????????????1233

Ala?Pro?Ser?Arg?Ser?Arg?Lys?Leu?Phe?Phe

405?????????????????410

<210>136

<211>410

<212>PRT

＜213〉marigold

<400>136

Met?Ala?Thr?His?Lys?Leu?Leu?Gln?Phe?Thr?Thr?Asn?Leu?Pro?Pro?Ser

1???????????????5???????????????????10??????????????????15

Ser?Ser?Ser?Ile?Ser?Thr?Gly?Cys?Ser?Leu?Ser?Pro?Phe?Phe?Leu?Lys

20??????????????????25??????????????????30

Ser?Ser?Ser?His?Ser?Pro?Asn?Pro?Arg?Arg?His?Arg?Arg?Ser?Ala?Val

35??????????????????40??????????????????45

Cys?Cys?Ser?Phe?Ala?Ser?Leu?Asp?Ser?Ala?Lys?Ile?Lys?Val?Val?Gly

50??????????????????55??????????????????60

Val?Gly?Gly?Gly?Gly?Asn?Asn?Ala?Val?Asn?Arg?Met?Ile?Gly?Ser?Gly

65??????????????????70??????????????????75??????????????????80

Leu?Gln?Gly?Val?Asp?Phe?Tyr?Ala?Ile?Asn?Thr?Asp?Ser?Gln?Ala?Leu

85??????????????????90??????????????????95

Leu?Gln?Ser?Val?Ala?His?Asn?Pro?Ile?Gln?Ile?Gly?Glu?Leu?Leu?Thr

100?????????????????105?????????????????110

Arg?Gly?Leu?Gly?Thr?Gly?Gly?Asn?Pro?Leu?Leu?Gly?Glu?Gln?Ala?Ala

115?????????????????120?????????????????125

Glu?Glu?Ser?Lys?Glu?Ala?Ile?Gly?Asn?Ala?Leu?Lys?Gly?Ser?Asp?Leu

130?????????????????135?????????????????140

Val?Phe?Ile?Thr?Ala?Gly?Met?Gly?Gly?Gly?Thr?Gly?Ser?Gly?Ala?Ala

145?????????????????150?????????????????155?????????????????160

Pro?Val?Val?Ala?Gln?Ile?Ala?Lys?Glu?Ala?Gly?Tyr?Leu?Thr?Val?Gly

165?????????????????170?????????????????175

Val?Val?Thr?Tyr?Pro?Phe?Ser?Phe?Glu?Gly?Arg?Lys?Arg?Ser?Val?Gln

180?????????????????185?????????????????190

Ala?Leu?Glu?Ala?Ile?Glu?Lys?Leu?Gln?Lys?Asn?Val?Asp?Thr?Leu?Ile

195?????????????????200?????????????????205

Val?Ile?Pro?Asn?Asp?Arg?Leu?Leu?Asp?Ile?Ala?Asp?Glu?Asn?Thr?Pro

210?????????????????215?????????????????220

Leu?Gln?Asp?Ala?Phe?Leu?Leu?Ala?Asp?Asp?Val?Leu?Arg?Gln?Gly?Val

225?????????????????230?????????????????235?????????????????240

Gln?Gly?Ile?Ser?Asp?Ile?Ile?Thr?Ile?Pro?Gly?Leu?Val?Asn?Val?Asp

245?????????????????250?????????????????255

Phe?Ala?Asp?Val?Lys?Ala?Val?Met?Lys?Asp?Ser?Gly?Thr?Ala?Met?Leu

260?????????????????265?????????????????270

Gly?Val?Gly?Val?Ser?Ser?Ser?Lys?Asn?Arg?Ala?Glu?Glu?Ala?Ala?Glu

275?????????????????280?????????????????285

Gln?Ala?Thr?Leu?Ala?Pro?Leu?Ile?Gly?Ser?Ser?Ile?Gln?Ser?Ala?Thr

290?????????????????295?????????????????300

Gly?Val?Val?Tyr?Asn?Ile?Thr?Gly?Gly?Lys?Asp?Ile?Thr?Leu?Gln?Glu

305?????????????????310?????????????????315?????????????????320

Val?Asn?Arg?Val?Ser?Gln?Val?Val?Thr?Ser?Leu?Ala?Asp?Pro?Ser?Ala

325?????????????????330?????????????????335

Asn?Ile?Ile?Phe?Gly?Ala?Val?Val?Asp?Glu?Arg?Tyr?Asn?Gly?Glu?Ile

340?????????????????345?????????????????350

His?Val?Thr?Ile?Val?Ala?Thr?Gly?Phe?Ala?Gln?Ser?Phe?Gln?Lys?Ser

355?????????????????360?????????????????365

Leu?Leu?Ala?Asp?Pro?Lys?Gly?Ala?Lys?Leu?Val?Asp?Arg?Asn?Gln?Glu

370?????????????????375?????????????????380

Pro?Thr?Gln?Pro?Leu?Thr?Ser?Ala?Arg?Ser?Leu?Thr?Thr?Pro?Ser?Pro

385?????????????????390?????????????????395?????????????????400

Ala?Pro?Ser?Arg?Ser?Arg?Lys?Leu?Phe?Phe

405?????????????????410

<210>137

<211>891

<212>DNA

＜213〉marigold

<220>

<221>CDS

<222>(1)..(891)

<223>

<400>137

atg?aca?tcc?ctg?agg?ttt?cta?aca?gaa?ccc?tca?ctt?gta?tgc?tca?tcc?????48

Met?Thr?Ser?Leu?Arg?Phe?Leu?Thr?Glu?Pro?Ser?Leu?Val?Cys?Ser?Ser

1???????????????5???????????????????10??????????????????15

act?ttc?ccc?aca?ttc?aat?ccc?cta?cac?aaa?acc?cta?act?aaa?cca?aca?????96

Thr?Phe?Pro?Thr?Phe?Asn?Pro?Leu?His?Lys?Thr?Leu?Thr?Lys?Pro?Thr

20??????????????????25??????????????????30

cca?aaa?ccc?tac?cca?aag?cca?cca?cca?att?cgc?tcc?gtc?ctt?caa?tac????144

Pro?Lys?Pro?Tyr?Pro?Lys?Pro?Pro?Pro?Ile?Arg?Ser?Val?Leu?Gln?Tyr

35??????????????????40??????????????????45

aat?cgc?aaa?cca?gag?ctc?gcc?gga?gac?act?cca?cga?gtc?gtc?gca?atc????192

Asn?Arg?Lys?Pro?Glu?Leu?Ala?Gly?Asp?Thr?Pro?Arg?Val?Val?Ala?Ile

50??????????????????55??????????????????60

gac?gcc?gac?gtt?ggt?cta?cgt?aac?ctc?gat?ctt?ctt?ctc?ggt?ctc?gaa????240

Asp?Ala?Asp?Val?Gly?Leu?Arg?Asn?Leu?Asp?Leu?Leu?Leu?Gly?Leu?Glu

65??????????????????70??????????????????75??????????????????80

aac?cgc?gtc?aat?tac?acc?gtc?gtt?gaa?gtt?ctc?aac?ggc?gat?tgc?aga????288

Asn?Arg?Val?Asn?Tyr?Thr?Val?Val?Glu?Val?Leu?Asn?Gly?Asp?Cys?Arg

85??????????????????90??????????????????95

ctc?gac?caa?gcc?cta?gtt?cgt?gat?aaa?cgc?tgg?tca?aat?ttc?gaa?ttg????336

Leu?Asp?Gln?Ala?Leu?Val?Arg?Asp?Lys?Arg?Trp?Ser?Asn?Phe?Glu?Leu

100?????????????????105?????????????????110

ctt?tgt?att?tca?aaa?cct?agg?tca?aaa?ttg?cct?tta?gga?ttt?ggg?gga????384

Leu?Cys?Ile?Ser?Lys?Pro?Arg?Ser?Lys?Leu?Pro?Leu?Gly?Phe?Gly?Gly

115?????????????????120?????????????????125

aaa?gct?tta?gtt?tgg?ctt?gat?gca?tta?aaa?gat?agg?caa?gaa?ggt?tgc????432

Lys?Ala?Leu?Val?Trp?Leu?Asp?Ala?Leu?Lys?Asp?Arg?Gln?Glu?Gly?Cys

130?????????????????135?????????????????140

ccg?gat?ttt?ata?ctt?ata?gat?tgt?cct?gca?ggt?att?gat?gcc?ggg?ttc????480

Pro?Asp?Phe?Ile?Leu?Ile?Asp?Cys?Pro?Ala?Gly?Ile?Asp?Ala?Gly?Phe

145?????????????????150?????????????????155?????????????????160

ata?acc?gcc?att?aca?ccg?gct?aac?gaa?gcc?gta?tta?gtt?aca?aca?cct????528

Ile?Thr?Ala?Ile?Thr?Pro?Ala?Asn?Glu?Ala?Val?Leu?Val?Thr?Thr?Pro

165?????????????????170?????????????????175

gat?att?act?gca?ttg?aga?gat?gca?gat?aga?gtt?aca?ggc?ttg?ctt?gaa????576

Asp?Ile?Thr?Ala?Leu?Arg?Asp?Ala?Asp?Arg?Val?Thr?Gly?Leu?Leu?Glu

180?????????????????185?????????????????190

tgt?gat?gga?att?agg?gat?att?aaa?atg?att?gtg?aac?aga?gtt?aga?act????624

Cys?Asp?Gly?Ile?Arg?Asp?Ile?Lys?Met?Ile?Val?Asn?Arg?Val?Arg?Thr

195?????????????????200?????????????????205

gat?ttg?ata?agg?ggt?gaa?gat?atg?atg?tca?gtt?ctt?gat?gtt?caa?gag????672

Asp?Leu?Ile?Arg?Gly?Glu?Asp?Met?Met?Ser?Val?Leu?Asp?Val?Gln?Glu

210?????????????????215?????????????????220

atg?ttg?gga?ttg?tca?ttg?ttg?agt?gat?acc?cga?gga?ttc?gaa?gtg?att????720

Met?Leu?Gly?Leu?Ser?Leu?Leu?Ser?Asp?Thr?Arg?Gly?Phe?Glu?Val?Ile

225?????????????????230?????????????????235?????????????????240

cgg?agt?acg?aat?aga?ggg?ttt?ccg?ctt?gtg?ttg?aac?aag?cct?ccg?act????768

Arg?Ser?Thr?Asn?Arg?Gly?Phe?Pro?Leu?Val?Leu?Asn?Lys?Pro?Pro?Thr

245?????????????????250?????????????????255

tta?gca?gga?ttg?gca?ttt?gag?cag?gct?gct?tgg?aga?ttg?gtt?gag?caa????816

Leu?Ala?Gly?Leu?Ala?Phe?Glu?Gln?Ala?Ala?Trp?Arg?Leu?Val?Glu?Gln

260?????????????????265?????????????????270

gat?agc?atg?aag?gct?gtg?atg?gtg?gag?gaa?gaa?cct?aaa?aag?agg?gga????864

Asp?Ser?Met?Lys?Ala?Val?Met?Val?Glu?Glu?Glu?Pro?Lys?Lys?Arg?Gly

275?????????????????280?????????????????285

ttt?ttc?tcg?ttt?ttt?gga?ggt?tag?tga????????????????????????????????891

Phe?Phe?Ser?Phe?Phe?Gly?Gly

290?????????????????295

<210>138

<211>295

<212>PRT

＜213〉marigold

<400>138

Met?Thr?Ser?Leu?Arg?Phe?Leu?Thr?Glu?Pro?Ser?Leu?Val?Cys?Ser?Ser

1???????????????5???????????????????10??????????????????15

Thr?Phe?Pro?Thr?Phe?Asn?Pro?Leu?His?Lys?Thr?Leu?Thr?Lys?Pro?Thr

20??????????????????25??????????????????30

Pro?Lys?Pro?Tyr?Pro?Lys?Pro?Pro?Pro?Ile?Arg?Ser?Val?Leu?Gln?Tyr

35??????????????????40??????????????????45

Asn?Arg?Lys?Pro?Glu?Leu?Ala?Gly?Asp?Thr?Pro?Arg?Val?Val?Ala?Ile

50??????????????????55??????????????????60

Asp?Ala?Asp?Val?Gly?Leu?Arg?Asn?Leu?Asp?Leu?Leu?Leu?Gly?Leu?Glu

65??????????????????70??????????????????75??????????????????80

Asn?Arg?Val?Asn?Tyr?Thr?Val?Val?Glu?Val?Leu?Asn?Gly?Asp?Cys?Arg

85??????????????????90??????????????????95

Leu?Asp?Gln?Ala?Leu?Val?Arg?Asp?Lys?Arg?Trp?Ser?Asn?Phe?Glu?Leu

100?????????????????105?????????????????110

Leu?Cys?Ile?Ser?Lys?Pro?Arg?Ser?Lys?Leu?Pro?Leu?Gly?Phe?Gly?Gly

115?????????????????120?????????????????125

Lys?Ala?Leu?Val?Trp?Leu?Asp?Ala?Leu?Lys?Asp?Arg?Gln?Glu?Gly?Cys

130?????????????????135?????????????????140

Pro?Asp?Phe?Ile?Leu?Ile?Asp?Cys?Pro?Ala?Gly?Ile?Asp?Ala?Gly?Phe

145?????????????????150?????????????????155?????????????????160

Ile?Thr?Ala?Ile?Thr?Pro?Ala?Asn?Glu?Ala?Val?Leu?Val?Thr?Thr?Pro

165?????????????????170?????????????????175

Asp?Ile?Thr?Ala?Leu?Arg?Asp?Ala?Asp?Arg?Val?Thr?Gly?Leu?Leu?Glu

180?????????????????185?????????????????190

Cys?Asp?Gly?Ile?Arg?Asp?Ile?Lys?Met?Ile?Val?Asn?Arg?Val?Arg?Thr

195?????????????????200?????????????????205

Asp?Leu?Ile?Arg?Gly?Glu?Asp?Met?Met?Ser?Val?Leu?Asp?Val?Gln?Glu

210?????????????????215?????????????????220

Met?Leu?Gly?Leu?Ser?Leu?Leu?Ser?Asp?Thr?Arg?Gly?Phe?Glu?Val?Ile

225?????????????????230?????????????????235?????????????????240

Arg?Ser?Thr?Asn?Arg?Gly?Phe?Pro?Leu?Val?Leu?Asn?Lys?Pro?Pro?Thr

245?????????????????250?????????????????255

Leu?Ala?Gly?Leu?Ala?Phe?Glu?Gln?Ala?Ala?Trp?Arg?Leu?Val?Glu?Gln

260?????????????????265?????????????????270

Asp?Ser?Met?Lys?Ala?Val?Met?Val?Glu?Glu?Glu?Pro?Lys?Lys?Arg?Gly

275?????????????????280?????????????????285

Phe?Phe?Ser?Phe?Phe?Gly?Gly

290?????????????????295

<210>139

<211>332

<212>DNA

＜213〉marigold

<220>

<221>CDS

<222>(1)..(330)

<223>

<400>139

aag?ctt?gca?cga?gcc?tct?ctc?tat?ttt?tac?act?tca?atg?gcg?gca?gca????48

Lys?Leu?Ala?Arg?Ala?Ser?Leu?Tyr?Phe?Tyr?Thr?Ser?Met?Ala?Ala?Ala

1???????????????5???????????????????10??????????????????15

att?gct?gtc?cct?tgt?agc?tca?aga?cca?ttt?ggc?tta?ggt?cga?atg?cgg????96

Ile?Ala?Val?Pro?Cys?Ser?Ser?Arg?Pro?Phe?Gly?Leu?Gly?Arg?Met?Arg

20??????????????????25??????????????????30

tta?ctt?ggt?cat?aaa?ccc?aca?acc?ata?act?tgt?cac?ttc?ccc?ttt?tct????144

Leu?Leu?Gly?His?Lys?Pro?Thr?Thr?Ile?Thr?Cys?His?Phe?Pro?Phe?Ser

35??????????????????40??????????????????45

ttt?tct?atc?aaa?tca?ttt?acc?cca?att?gtt?agg?ggc?aga?aga?tgt?act????192

Phe?Ser?Ile?Lys?Ser?Phe?Thr?Pro?Ile?Val?Arg?Gly?Arg?Arg?Cys?Thr

50??????????????????55??????????????????60

gtt?tgt?ttt?gtt?gcc?ggt?ggc?gac?agt?aat?agt?aac?agt?aat?aat?aat????240

Val?Cys?Phe?Val?Ala?Gly?Gly?Asp?Ser?Asn?Ser?Asn?Ser?Asn?Asn?Asn

65??????????????????70??????????????????75??????????????????80

agt?gac?agt?aat?agt?aat?aat?ccg?ggt?ctg?gat?tta?aac?ccg?gcg?gtt????288

Ser?Asp?Ser?Asn?Ser?Asn?Asn?Pro?Gly?Leu?Asp?Leu?Asn?Pro?Ala?Val

85??????????????????90??????????????????95

atg?aac?cgt?aac?cgt?ttg?gtt?gaa?gaa?aaa?atg?gag?agg?tcg?ac?????????332

Met?Asn?Arg?Asn?Arg?Leu?Val?Glu?Glu?Lys?Met?Glu?Arg?Ser

100?????????????????105?????????????????110

<210>140

<211>110

<212>PRT

＜213〉marigold

<400>140

Lys?Leu?Ala?Arg?Ala?Ser?Leu?Tyr?Phe?Tyr?Thr?Ser?Met?Ala?Ala?Ala

1???????????????5???????????????????10??????????????????15

Ile?Ala?Val?Pro?Cys?Ser?Ser?Arg?Pro?Phe?Gly?Leu?Gly?Arg?Met?Arg

20??????????????????25??????????????????30

Leu?Leu?Gly?His?Lys?Pro?Thr?Thr?Ile?Thr?Cys?His?Phe?Pro?Phe?Ser

35??????????????????40??????????????????45

Phe?Ser?Ile?Lys?Ser?Phe?Thr?Pro?Ile?Val?Arg?Gly?Arg?Arg?Cys?Thr

50??????????????????55??????????????????60

Val?Cys?Phe?Val?Ala?Gly?Gly?Asp?Ser?Asn?Ser?Asn?Ser?Asn?Asn?Asn

65??????????????????70??????????????????75??????????????????80

Ser?Asp?Ser?Asn?Ser?Asn?Asn?Pro?Gly?Leu?Asp?Leu?Asn?Pro?Ala?Val

85??????????????????90??????????????????95

Met?Asn?Arg?Asn?Arg?Leu?Val?Glu?Glu?Lys?Met?Glu?Arg?Ser

100?????????????????105?????????????????110

<210>141

<211>332

<212>DNA

＜213〉marigold

<220>

<221>misc_feature

<222>(1)..(332)

＜223〉B-hydroxylase is had a mind to fragment

<400>141

aagcttgcac?gagcctctct?ctatttttac?acttcaatgg?cggcagcaat?tgctgtccct?????60

tgtagctcaa?gaccatttgg?cttaggtcga?atgcggttac?ttggtcataa?acccacaacc????120

ataacttgtc?acttcccctt?ttctttttct?atcaaatcat?ttaccccaat?tgttaggggc????180

agaagatgta?ctgtttgttt?tgttgccggt?ggcgacagta?atagtaacag?taataataat????240

agtgacagta?atagtaataa?tccgggtctg?gatttaaacc?cggcggttat?gaaccgtaac????300

cgtttggttg?aagaaaaaat?ggagaggtcg?ac??????????????????????????????????332

<210>142

<211>332

<212>DNA

＜213〉marigold

<220>

<221>misc_feature

<222>(1)..(332)

＜223〉B-hydroxylase antisense fragment

<400>142

gaattcggca?cgagcctctc?tctattttta?cacttcaatg?gcggcagcaa?ttgctgtccc?????60

ttgtagctca?agaccatttg?gcttaggtcg?aatgcggtta?cttggtcata?aacccacaac????120

cataacttgt?cacttcccct?tttctttttc?tatcaaatca?tttaccccaa?ttgttagggg????180

cagaagatgt?actgtttgtt?ttgttgccgg?tggcgacagt?aatagtaaca?gtaataataa????240

tagtgacagt?aatagtaata?atccgggtct?ggatttaaac?ccggcggtta?tgaaccgtaa????300

ccgtttggtt?gaagaaaaaa?tggagaggat?cc??????????????????????????????????332

Claims

1. the purposes of the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part is used to be administered orally in animal.

2. purposes according to claim 1, wherein the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part is used for painted animal and corresponding animal product.

3. purposes according to claim 1 and 2, wherein the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part mixes with the animal feed goods and these animal feed goods are administered orally in animal.

4. purposes according to claim 3, wherein with before the animal feed goods mix, the plants of tagetes species of astaxanthin-containing or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part are processed into the form that can mix with the animal feed goods.

5. purposes according to claim 1 and 2, wherein the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part directly is administered orally in animal.

6. purposes according to claim 5, wherein before using, the plants of tagetes species of astaxanthin-containing or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part are processed into the form that can directly be administered orally in animal.

7. according to any described purposes among the claim 1-6, wherein made the plants of tagetes species of astaxanthin-containing can produce astaxanthin by genetic manipulation.

8. according to any described purposes among the claim 1-7, wherein animal is selected from fish, crustacean, Galliformes and Anatridae.

9. purposes according to claim 8, wherein animal is selected from salmonid, shrimp, crab, chicken, duck, goose and flamingo.

10. according to any described purposes among the claim 2-9, wherein animal product is selected from meat, skin, feather and yolk.

11. according to any described purposes among the claim 1-10, wherein employed plant part is capitulum or petal.

12. produce the method for animal feed goods, it is with the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part and the combination of conventional animal feed ingredient.

13. method according to claim 12, wherein with animal feed combination before, with the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part be processed into can with the form of animal feed combination.

14. be used for the method for painted animal or animal product, it is administered orally in animal by the astaxanthin-containing extract with the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part and carries out.

15. be used to produce the painted animal or the method for animal product, it is for being administered orally in animal with the plants of tagetes species of astaxanthin-containing or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part.

16., wherein the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part is mixed with the animal feed goods and the animal feed goods is administered orally in animal according to claim 14 or 15 described methods.

17. method according to claim 16, wherein with before the animal feed goods mix, the plants of tagetes species of astaxanthin-containing or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part are processed into the form that can mix with the animal feed goods.

18. according to claim 14 or 15 described methods, wherein the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part directly is administered orally in animal.

19. method according to claim 18, before wherein using, the plants of tagetes species of astaxanthin-containing or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part are processed into the form that can directly be administered orally in animal.

20., wherein made the plants of tagetes species of astaxanthin-containing can produce astaxanthin by genetic manipulation according to any described method among the claim 14-19.

21. according to any described method among the claim 14-20, wherein animal is selected from fish, crustacean, Galliformes and Anatridae.

22. method according to claim 21, wherein animal is selected from salmonid, shrimp, crab, chicken, duck, goose and flamingo.

23. according to any described method among the claim 14-22, wherein animal product is selected from meat, skin, feather and ovum.

24. according to any described method among the claim 14-23, wherein employed plant part is capitulum or petal.

25. the purposes of the astaxanthin-containing extract of the plants of tagetes species of the plants of tagetes species of astaxanthin-containing or plant part or astaxanthin-containing or plant part is as animal feed or animal feed additive.

26. the animal feed goods, it comprises plants of tagetes species or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part of astaxanthin-containing.

27. colouring agent, it comprises plants of tagetes species or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part of astaxanthin-containing.

28. colouring agent according to claim 27, it is made up of the plants of tagetes species of astaxanthin-containing or the plants of tagetes species of plant part or astaxanthin-containing or the astaxanthin-containing extract of plant part.

29. according to claim 27 or 28 described colouring agents, wherein employed plant part is capitulum or petal.