CN101495626A - Identification of a novel type of sucrose synthase and use thereof in fiber modification - Google Patents

Identification of a novel type of sucrose synthase and use thereof in fiber modification Download PDF

Info

Publication number
CN101495626A
CN101495626A CNA2007800286149A CN200780028614A CN101495626A CN 101495626 A CN101495626 A CN 101495626A CN A2007800286149 A CNA2007800286149 A CN A2007800286149A CN 200780028614 A CN200780028614 A CN 200780028614A CN 101495626 A CN101495626 A CN 101495626A
Authority
CN
China
Prior art keywords
leu
glu
val
lys
gly
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CNA2007800286149A
Other languages
Chinese (zh)
Inventor
阮永良
R·富尔班克
E·布里尔
M·范托尔诺特
A·阿廖利
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Commonwealth Scientific and Industrial Research Organization CSIRO
Bayer CropScience NV
Original Assignee
Commonwealth Scientific and Industrial Research Organization CSIRO
Bayer CropScience NV
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Commonwealth Scientific and Industrial Research Organization CSIRO, Bayer CropScience NV filed Critical Commonwealth Scientific and Industrial Research Organization CSIRO
Publication of CN101495626A publication Critical patent/CN101495626A/en
Pending legal-status Critical Current

Links

Images

Abstract

Methods and means are described to modify fiber characteristics in a fiber producing plant, such as cotton, based on the use of a novel type of sucrose synthase protein or related nucleic acids.

Description

The evaluation of novel type of sucrose synthase and the purposes in fibre modification thereof
Technical field
The present invention relates to agriculture field, relate more specifically to Protocols in Molecular Biology and be used to change the fiber production plant, especially vegetable lamb and/or quicken the purposes of the breeding of this type of fibrous plants.The invention provides, for example, change the ways and means of fiber quality by increasing or reduce the grand Buddhist nun's value of content of cellulose, fibre strength, fibre uniformity or mark (micronaire).Also provide and in cotton variety colony and relevant ancestors plant, identified the relevant molecular marker method of category feature therewith.
Background technology
The many high product amount fiber that uses in the textile industry is provided by cotton.About 90% is upland cotton (Gossypium hirsutum L.) in the cotton of whole world plantation, and sea island cotton (Gossypiumbarbadense) accounts for about 8%.Therefore, in the breeding of being undertaken by classical way or the genome by the hereditary change vegetable lamb, mainly making great efforts is to transform the cotton fiber feature to adapt to the industry needs better.The target that realizes comprises velveteen (lint) staple length, intensity, the dyeability of increase, the grand Buddhist nun's value of linters (fuzz fiber) generation, fiber maturity ratio (fiber maturity ratio), immature fibre content, fibre uniformity and mark of minimizing.
US6472588 and WO0117333 provide by the DNA with the encoding sucrose phosphate synthase and have transformed, and increase from the method for quality of the cotton fibre of vegetable lamb generation.This fiber quality comprises that intensity, length, fiber maturity ratio, immature fibre content, fibre uniformity and mark swell Buddhist nun's value.
WO9508914 discloses the fiber production plant that comprises the allogeneic heredity construct in its genome.This genetic constructs comprises fiber-specific promotor and coded plant peroxidase, for example cotton peroxidase, encoding sequence.
WO9626639 provides following method, wherein utilizes preferentially in the ovary tissue, and particularly extremely early stage at fruit development instructs the encoding sequence of genetic expression to express the plant growth regulating hormone in cotton ovule tissue.This method allows to change knot bell (boll set) feature of cotton plant and provides and changes the fiber quality feature, for example fiber size and intensity, mechanism.
US5981834, US5597718, US5620882, US5521708 and US 5495070 all disclose and have been used for genetic engineering modified fiber production plant and identify the method for cloning at the useful cDNA of the evaluation of cotton fiber gene.These cDNA clones can be used for developing corresponding genomic clone from the fiber production plant, so that can utilize these gene genetics to transform cotton and other plant.Encoding sequence from these isolating genes can use so that justice or antisense orientation to be arranged, thereby changes transgenosis fiber production fibre of plant feature.
Gossypium (Gossypium) plant that U.S. Patent application US2002049999 that announces and US2003074697 disclose the cotton fibre feature with improvement.This vegetable lamb has the expression of gene box of the enzyme that comprising encodes is selected from endoxyloglucan transferring enzyme, catalase and peroxidase, and this gene is expressed in cotton fiber cell thus, thus improvement cotton fibre feature.
US5880110 handles by using the plain steroid of rape, produces the cotton fibre of the fiber characteristics with improvement.
WO01/40250 is provided for expressing the method that improves cotton quality by regulating transcription factor gene.
WO96/40924 provides the Novel DNA construct, and described construct can be used as molecular probe or the insertion plant host changes transcribing of target DNA sequence with the different steps at cotton fiber development.This DNA construct comprises the cotton fibre transcription initiation control region that is connected with the gene for the treatment of to express in cotton fibre.By using this construct in cotton fiber cell, to express the cotton of chromogene cotton fibre that produce, that have natural colour, also be new.
EP0834566 is provided at the gene that controlling fiber in the cotton plant forms mechanism and can be used for industrial useful improvement.
(The Plant Cell, vol 15,952-964) described initial, extension and seed development that the inhibition that sucrose synthase gene is expressed can be prevented cotton fiber cell for people such as Ruan 2003.Result described herein provides the expression of sucrose synthase in the ovule epidermis to extend vital direct evidence for the initial sum of unicellular fibre.Can also draw by described result: in kind of skin, suppress Sus and only reduce the growth of fiber and do not influence fetal development and seed size.The document draws as drawing a conclusion: will provide by genetic modification Sus about the new knowledge of the effect of Sus in controlling plant cell and seed development and express the chance that changes fiber and seed development.
WO0245485 describes by regulating the active of sucrose synthase and/or express in the fiber production plant and changes for example ways and means of the fiber quality in the cotton of this type of plant.
People such as Baud 2004 (Journal of Experimental Botany, the 55th volume, No.396, pp 397-409) have described the structure and the express spectra that have 6 members' sucrose synthase multigene family in the Arabidopsis (Arabidopsis).
None described the identification to such sucrose synthase of recognizing at present in the prior art document, did not also describe its (for example C type) and participate in secondary plant cell wall growth in plant.This New type of S us albumen be provided for changing the fiber production plant for example the new and improved method and the means of the fiber characteristics of cotton opened possibility, these ways and meanses and any other method that changes fiber characteristics further can be made up.This type of ways and means has been described in embodiment hereinafter and the claim.
Summary of the invention
In one embodiment of the invention, novel type of sucrose synthase with following aminoacid sequence albumen is provided, and this aminoacid sequence comprises and is selected from following aminoacid sequence: have the aminoacid sequence of at least 50% sequence homology with SEQ ID Nos.:2, any aminoacid sequence of 3,5,7,9,11 or 13; Comprise SEQ ID Nos.:2, any aminoacid sequence of aminoacid sequence of 3,5,7,9,11 or 13; Be positioned at the aminoacid sequence of described proteinic aminoterminal part, described aminoacid sequence comprises the sequence identity with the aminoacid sequence about at least 60% of SEQ ID No.16; Or being positioned at the aminoacid sequence of described proteinic carboxyl terminal part, described aminoacid sequence comprises the sequence identity with the aminoacid sequence about at least 60% of SEQ ID No.15.Described sucrose synthase albumen can comprise hydrophobicity N terminal sequence.It also can comprise the arbitrary of following amino acid sequences: 394 the aminoacid sequence from amino acid 383 to amino acid of SEQ IDNo.:3; 329 the aminoacid sequence of SEQ ID No.:3 from amino acid 270 to amino acid; 737 the aminoacid sequence of SEQ ID No.:3 from amino acid 549 to amino acid; 545 the aminoacid sequence of SEQ ID No.:3 from amino acid/11 to amino acid; Or 794 the aminoacid sequence of SEQ ID No.:3 from amino acid/11 8 to amino acid.
In another embodiment of the invention, provide the isolating novel type of sucrose synthase of identification proteic antibody.
In another embodiment, the invention provides proteic isolated DNA molecule of encoding novel sucrose synthase or nucleic acid, for example comprise the nucleic acid of SEQ ID Nos.:1, arbitrary nucleotide sequence of 4,6,8,10,12 or 14.
The present invention also provides the expression cassette of the dna molecular that comprises following effective connection: plant can be expressed promotor, and for example preferential plant that control is transcribed in fiber production fibre of plant cell can be expressed promotor; The proteic DNA of novel type of sucrose synthase that coding is provided; Randomly Transcription Termination and polyadenylation region.
The present invention also provides the expression cassette of the dna molecular that comprises following effective connection: plant can be expressed promotor; Produce the DNA of RNA molecule when transcribing, wherein said RNA molecule comprises with the nucleotide sequence of endogenous sucrose synthase C isotype (isoform) encoding gene or with the complementary sequence of the nucleotide sequence of described endogenous sucrose synthase C isotype encoding gene and has the nucleotide sequence of at least 19 continuous nucleotides of about at least 94% sequence identity, or comprises the nucleotide sequence that has at least 19 continuous nucleotides of about at least 94% sequence identity with the nucleotide sequence of novel C isotype sucrose synthase; Randomly Transcription Termination and polyadenylation region.
Another embodiment of the invention is the expression cassette that comprises the dna molecular of following effective connection: plant can be expressed promotor, and the preferred preferential plant that control is transcribed in fibrocyte can be expressed promotor; Transcribe the DNA that produces double stranded rna molecule, described RNA molecule can reduce the endogenous SusC genetic expression of fiber production plant, and described RNA molecule comprises the first and second RNA zones, and wherein a RNA zone comprises with the nucleotide sequence of endogenous SusC gene or with the proteic nucleotide sequence of coding SusC that provides and has the nucleotide sequence of at least 19 continuous nucleotides of about at least 94% sequence identity; The 2nd RNA zone comprises these at least 19 the continuous nucleotide complementary nucleotide sequences with a RNA zone; The first and second RNA zones can base pairing, thereby forms double stranded rna molecule between these at least 19 continuous nucleotides in first and second zones; Randomly be included in the 3 ' stub area that has the Transcription Termination and the polyadenylation signal of function in the cell of this plant.
The present invention also provides and comprises the vegetable cell that the allos plant that effectively is connected with the dna molecular that is selected from following dna molecular can be expressed promotor: the DNA of the inhibitory RNA of the DNA of the C isotype sucrose synthase of encoding new or coding susC encoding gene.This cell can be from the fiber production plant cell of cotton for example.The present invention also comprises the fiber that comprises this type of vegetable cell or plant, seed or the plant part be made up of this class vegetable cell basically or tissue and produced by this class plant.
In another embodiment of the invention, be provided for the method for fibres modified production fibre of plant feature, described method is included in cell or cell walls or the middle change of apoplast liquid (apoplastic fluid) of described plant, for example increase or reduce, sucrose synthase C isotype or have the functional level of the sucrose synthase of similar features.This can realize easily by expression cassette described herein is provided to plant.
The present invention comprises that also new SusC type albumen or coding nucleic acid are used for changing the fiber production plant purposes of fiber characteristics.
The accompanying drawing summary
Fig. 1 .SusC albumen and the proteic hydrophobicity collection of illustrative plates of similar sucrose.A. from the collection of illustrative plates of the SusC of upland cotton kind; B: from the collection of illustrative plates of the SusC of sea island cotton kind; C: from the collection of illustrative plates of the SusC of tree cotton (Gossypium arboretum); D: from the collection of illustrative plates of the SusC of Lei Mengdeshi cotton (Gossypiumraimondii); E: from the collection of illustrative plates of the SusC of long calyx cotton (Gossypium longicalyx) hypotype 1; F: from the collection of illustrative plates of the SusC of the cotton hypotype 2 of long calyx; G: from the collection of illustrative plates of the sucrose synthase of mung bean (Vigna radiata); H: the collection of illustrative plates that comes the sucrose synthase of arrogant eucalyptus (Eucalyptus grandis); I: from the collection of illustrative plates of the sucrose synthase isotype 6 of Arabidopis thaliana (Arabidopsis thaliana); K: from the collection of illustrative plates of the sucrose synthase of Populus tremuloides (Populus tremuloides).All hydrophobicity collection of illustrative plates have all shown the hydrophobicity N end of these different sucrose synthases.
Fig. 2. the SusC expression of gene analysis of in cotton fiber tissue, carrying out by RT-PCR.Dpa: the back fate of blooming; GDNA: crt gene group DNA.Swimming lane 1 and 8:1kb mark.
Fig. 3: from intron and the exon in the sucrose synthase encoding gene of cotton.The intron of diagram C type sucrose synthase distributes with exon, and with Type B and the comparison of A type sucrose synthase from cotton.Black surround is represented exon.
Fig. 4: from the 2D structural analysis of the N end parts of the sucrose synthase gene of cotton.Compare the 2D structural analysis (HCA+JPRED) of the N end parts of susyC gene with other upland cotton sucrose synthases.Arrow shows that this susyC specific regions is to the influence of 2D-structure on every side.(P)=phosphorylation site inferred.
Fig. 5: from the 2D structural analysis of the C end parts of the sucrose synthase gene of cotton.In this susyC 2D structure, as if there is not the βZhe Die chip architecture.
Fig. 6: the phenotype analytical of transgenic arabidopsis platymiscium.Fasciation (fasciated) stem of demonstration chap bifurcated in 2 independent transgenic lines with CaMV 35S-SusC mosaic gene (figure A and B).The T2 of figure C and D demonstration CaMV 35S-SusC and S2A-SusC mosaic gene typical trichome phenotype in generation.Figure C is three-dimensional photomontage (photo-montage).
Fig. 7: use SUS C specific antibody to having the Western trace of the genetically modified T2 plantlet of CaMV 35S-SusC.Swimming lane A: plant with slight phenotype; Swimming lane B: have the plant of severe phenotype, swimming lane C: plant with slight phenotype; Swimming lane D: plant with severe phenotype; WT: wild-type plant.
Detailed Description Of The Invention
The present invention is based on the evaluation from the new sucrose synthase isotype (being called SusC or SuSyC) of fiber, the express spectra (expression profile) of described sucrose synthase isotype on rna level and procedure of fibre production plant for example in the cotton secondary cell wall budding initial closely related. This protein is very abundant in the secondary cell wall forming process, mainly is arranged in apoplast. Its location, abundance and express spectra show that this novel type of sucrose synthase isotype is to be present in main isotype in the cell membrane at the secondary cell wall synthesis phase. Be not intended to limit the invention to specific binding mode, this sucrose synthase isotype is considered to participate in from picking sugar and incorporate them into cellulose and/or callose with UDP glucose form around the apoplast liquid of fiber.
Therefore, in the first embodiment, the invention provides the method that changes the cell membrane feature in plant cell, described method is characterised in that this new sucrose synthase (SusC) isotype of change or has the functional level of the sucrose synthase of similar features.
Sucrose synthase C type isotype or the functional level with sucrose synthase of similar features can change by the expression that increases this type of isotype in plant cell. This can comprise by importing the nucleic acid of following effective connection, for example the expression construct of dna molecular, easily realization:
A) plant can be expressed promoter
B) encoding sucrose synthase C isotype or have the nucleic acid of the sucrose synthase of similar features; Randomly
C) tanscription termination and polyadenylation region.
As used herein, " sucrose synthase " refer to can catalysis from the enzyme of NDP-glucose (for example uridine diphosphoglucose) and D-Fructose synthesis of sucrose. But this enzyme also catalysis sucrose hydrolysis becomes glucose and fructose.
This enzyme classification is EC 2.4.1.13. The another name of sucrose synthase is glucosyltransferase, uridine diphosphoglucose-fructose, sucrose synthase, sucrose-UDP glucosyltransferase, sucrose-uridine diphosphate glucose based transferase, Sus, SuSy, UDPG-fructose glucosyltransferase, UDPG: D-Fructose 2-α-D-glucosyltransferase and uridine diphosphoglucose-fructose glucosyltransferase.
Several genes from the encoding sucrose synthase of plant have been described. WO02/45485 (the 16th to 20 page) provides the nucleotide sequence, its part of coded plant sucrose synthase gene or has had the full list of Genbank accession number of the nucleotide sequence of sequence similarity with sucrose synthase gene.
Described the active enzymatic determination method of sucrose synthase (referring to for example Counce and Gravois, 2006, Crop Science the 46th volume: pp1501-1507, special the 1502nd and 1503 page; Anthon and Emerich, 1990, the 92 volumes: pp 346-351, the 347th page especially; Two pieces of documents are integrated with this paper by reference).
" sucrose synthase C isotype " or " SusC " or " SuSyC " are characterised in that exist (referring to Fig. 1) of hydrophobicity N terminal amino acid sequence.Preceding 44 amino acid form hydrophobic region, particularly the residue 26 to 44 of SEQID No.:3 for example.The other plant sucrose synthase that is characterised in that this hydrophobicity N terminal amino acid sequence is the sucrose synthase 6 (accession number At1g73370) from Arabidopis thaliana; From the sucrose synthase (accession number D10266) of mung bean, belong to the sucrose synthase (accession number DD014303) of (Eucalyptus spp) and from the sucrose synthase (accession number AY341026) (all sequences is incorporated this paper by reference into) of willow from eucalyptus.
The comparison of different SusC aminoacid sequences and with the relatively demonstration from other sucrose synthases of plant exist the fragment of 20-25 the continuous amino acid that lays respectively at N end (SEQ ID No:16) or C end (SEQ ID No.15), described fragment in the SusC aminoacid sequence be guard and be not present in the other plant sucrose synthase.N terminal sequence (HKSQKLLSVLDKEAGNQALDGMVV; SEQ ID No.:16) be usually located between amino acid position 36 and the amino acid position 59, and C terminal sequence (AYQEQRGRKRYIEMLHAWMYNNRVKT; SEQ ID No.:15) is usually located between amino acid position 765 and 790.
Actin muscle calmodulin binding domain CaM (people such as Winter H., 1998, FEBS letters 430,205-208 that sucrose synthase albumen has supposition have also been described; Winter H. and Huber S.C., 2000CriticalReviews in Plant Sciences 19 (1), 31-67), have following consensus amino acid sequences: KDVAAE[V/I at Actin muscle calmodulin binding domain CaM described in the SusC isotype] TKEFQ (amino acid position 383 of SEQ ID No 3 is to amino acid position 394).Usually the phenylalanine residue (F) that sees the lysine residue (K) of position 383 and see position 393 usually also is the proteic index of SusC type.
Also described the uridine calmodulin binding domain CaM that sucrose synthase albumen has supposition, had following amino acid sequences VVIMTPHGYFAQDNVLGYPDTGGQVVYILDQVRALEEELLHRFKLQGLDITPRILV ITRL (amino acid position 270 of SEQ ID No 3 is to amino acid position 329) at uridine calmodulin binding domain CaM described in the SusC isotype.
By using the computer based search, can in the aminoacid sequence of SusC isotype, identify following structural domain or sign.These structural domains or sign are common with other sucrose synthases from plant.
Glycosyltransferase is organized 1 structural domain: (IPR001296; PF00534)
The protein that comprises this structural domain is transferred to various substrates with the sugar that UDP, ADP, GDP or CMP connect, and comprises glycogen, fructose-6-phosphate and lipopolysaccharides.These bacterial enzymes participate in multiple biosynthetic process, comprise that exocellular polysaccharide biosynthesizing, the biosynthesizing of lipopolysaccharides core and slime polysaccharide can draw the biosynthesizing of acid.This structural domain corresponding to SEQ ID No 3 for example from the aminoacid sequence of amino acid position 549 to 737..
Sign (the IPR000368 of sucrose synthase family; PF00862):
This sign can characterize the family that comprises large quantities of sucrose synthase albumen.Yet the carboxyl terminal zone of these sucrose synthases belongs to glycosyltransferase family.This enzyme is mainly seen in the plant but also appears in the bacterium.This structural domain corresponding to SEQ ID No 3 for example from the aminoacid sequence of amino acid position 1 to 545.
Sucrose synthase, plant and cyanobacteria family sign (IPR012820)
This sign represent sucrose synthase, a kind of common use but not produce the enzyme (although it is called sucrose synthase) of sucrose.Sucrose+UDP (or ADP) forms D-fructose+UDP-glucose (or ADP-glucose), and it can be used for the biosynthesizing of cell walls (or starch) then.The sucrose phosphosynthase homology of the step second from the bottom during this enzyme and catalysis sucrose are synthetic.Up to the present, only in plant and cyanobacteria, find sucrose synthase.This structural domain is corresponding to for example from the aminoacid sequence of the SEQ ID No 3 of amino acid position 18 to 794.
Use PROSITE spectrum (can obtain PROSITE),, can easily discern these structural domains by the computer based search at www.expasy.ch/prosite.Selectively, also can use BLOCKS database and algorithm (blocks.fhcrc.org) to identify these structural domains.Also can obtain other databases and algorithm (pFAM:http: //www.sanger.ac.uk/Software/Pfam/; INTERPRO: Http:// www.ebi.ac.uk/interpro/Relate to pFAM database and algorithm above-cited PF number, relate to INTERPRO database and algorithm IPR number).
The invention provides variant aminoacid sequence and the nucleotide sequence (cDNA and genomic dna) of a large amount of separation from the SusC isotype of upland cotton Cultivar (SEQ ID Nos:3-4), sea island cotton Cultivar (SEQ ID Nos:5 and 6), tree cotton (SEQ ID Nos:7 and 8), Lei Mengdeshi cotton (SEQID Nos:9 and 10) and long calyx cotton (SEQ ID Nos.:11 to 14).
Other variant nucleotide sequences can be available from other upland cotton or sea island cotton kind or available from cotton ancestors plant tree cotton for example, cotton and the long calyx cotton of cotton (Gossypium herbaceum) and Lei Mengdeshi, or other Gossypium species, particularly comprise the genomic Gossypium species of D type, for example nonirrigated farmland cotton (Gossypium aridum), Dai Weixunshi cotton (Gossypium davidsonii), intend like cotton (Gossypium gossypioides), Ke Laociji cotton (Gossypium klotzschianum), plucked instrument Bai Shi cotton (Gossypium turberi), three split cotton (Gossypium trilobum), Te Nashi cotton (Gossypium turneri), yellowish-brown cotton (Gossypium mustelinum), Hawaii cotton (Gossypium tomentosum), Da Erwenshi cotton (Gossypium darwinii).Variant aminoacid sequence or nucleotide sequence comprise the change of the sequence of interpolation, disappearance or the displacement generation of passing through amino acid or Nucleotide respectively.Can be from comprising the nucleic acid of genomic other Gossypium species pcr amplification codings of D type SusC.The nucleic acid of this type of pcr amplification demonstrates and comprises the distinctive EcoRI restriction site of SusC type coding nucleic acid.
Can find the variant of sucrose synthase C isotype by strict hybridization, described hybridization use SEQID No 1,4,6,8,10,12 or 14 any nucleotide sequence or described sequence in comprise the part of about at least 25 or 50 continuous nucleotides in SEQ ID No SEQ ID No 1,4,6,8,10,12 or 14 or its complementary nucleotide sequence as probe.Useful especially probe can be the N end of coding SEQ ID 15 and 16 or the nucleotide sequence of C terminal sequence, for example, SEQID No 1,4,6,8,10,12 or 14 any in the nucleotide sequence of coding SEQ ID Nos 15 or 16 aminoacid sequence.
" stringent hybridization condition " as used herein is meant if having 95% between probe and the target sequence at least, preferred at least 97% sequence identity, and then hybridization will take place usually.The example of stringent hybridization condition is: contain 50% methane amide, 5x SSC (150mM NaCl, 15mM trisodium citrate), 50mM sodium phosphate (pH7.6), 5x Denhardt solution, 10% T 500 and 20 μ g/ml sex change, carrier DNA through shearing for example is incubated overnight in the solution of salmon sperm DNA, at about 65 ℃ of washing hybridization upholders in 0.1x SSC, preferably carried out about 10 minutes then.Other hybridization and wash conditions are known, people such as Sambrook, and Molecular Cloning:A LaboratoryManual, the 2nd edition, Cold Spring Harbor, NY (1989) illustrates it in the Chapter 11 especially.
Also can use for sucrose synthase (SusC) gene is that specific oligonucleotide is as primer, obtain this type of variant sequence by DNA cloning, described oligonucleotide is, such as but not limited to, by about 20 oligonucleotide or oligonucleotide that comprises it or their complementary sequences of forming to about 50 continuous nucleotides in the nucleotide sequence of SEQID No 1,4,6,8,10,12 or 14.SEQ IDNos 17 and 18 examples be especially suitable for use as the primer nucleotide sequence of SusC Auele Specific Primer.
Except the natural variation that utilizes the SusC isotype, also can obtain the variant sequence by inducing the generation variation in external or the body.The several method that is used for external evoked generation variant nucleotide sequence is obtainable in this area, includes but not limited to DNA reorganization or the orthogenesis technology described in United States Patent (USP) 5605793, United States Patent (USP) 5,811,238 and the United States Patent (USP) 5,830,721.Be used for inducing in the body method that produces the variant nucleotide sequence in this area, also to know, it can comprise vegetable lamb or cotton ancestors plant are exposed to for example ionizing rays of mutagenic compound, EMS, MMS etc., for example described then as elsewhere in this specification sheets, the nucleic acid of separation coding SusC.
Behind C end that identifies SusC type sucrose synthase and N end consensus sequence, the inventor has identified the following nucleotide sequences of the aminoacid sequence that comprises this consensus sequence of encoding in database.By using the N total zone of end (SEQ ID No.:16), separated following sequence (by their accession number sign): EMBL:CO499214 (upland cotton cDNA), EMBL:CO499090 (upland cotton cDNA), EMBL:CO498472 (upland cotton cDNA), EMBL:CO498387 (upland cotton cDNA), EMBL:CO497432 (upland cotton cDNA), EMBL:CO497275 (upland cotton cDNA), EMBL:CO496887 (upland cotton cDNA), EMBL:CO496593 (upland cotton cDNA), EMBL:CO495954 (upland cotton cDNA), EMBL:CO495804 (upland cotton cDNA), EMBL:CO495643 (upland cotton cDNA), EMBL:CO495623 (upland cotton cDNA), EMBL:CO495601 (upland cotton cDNA), EMBL:CO495511 (upland cotton cDNA), EMBL:CO494831 (upland cotton cDNA), EMBL:CO494779 (upland cotton cDNA), EMBL:CO494693 (upland cotton cDNA), EMBL:CO494536 (upland cotton cDNA), EMBL:CO494182 (upland cotton cDNA), EMBL:CO493935 (upland cotton cDNA), EMBL:CO493780 (upland cotton cDNA), EMBL:CO493725 (upland cotton cDNA), EMBL:CO493724 (upland cotton cDNA), EMBL:CO493589 (upland cotton cDNA), EMBL:CO493043 (upland cotton cDNA), EMBL:CO493007 (upland cotton cDNA), EMBL:CO492969 (upland cotton cDNA), EMBL:CO492566 (upland cotton cDNA), EMBL:CO490959 (upland cotton cDNA), EMBL:BF272227 (tree cotton cDNA clones GA__Eb0014E15f), EMBL:CO493732 (upland cotton cDNA), EMBL:CO493111 (upland cotton cDNA), EMBL:CO491540 (upland cotton cDNA), EMBL:CO493583 (upland cotton cDNA), EMBL:CO493765 (upland cotton cDNA) and EMBL:CO498054 (upland cotton cDNA).The sequence of all citations is integrated with this paper by reference.By using C end consensus sequence (SEQ ID No.:15), separated following sequence (by their accession number sign): EMBL:DT463218 (upland cotton cDNA clones GH_CHX13D22-3), EMBL:CO499051 (upland cotton cDNA), EMBL:CO498850 (upland cotton cDNA), EMBL:CO498685 (upland cotton cDNA), EMBL:CO498361 (upland cotton cDNA), EMBL:CO498044 (upland cotton cDNA), EMBL:CO497702 (upland cotton cDNA), EMBL:CO497506 (upland cotton cDNA), EMBL:CO497291 (upland cotton cDNA), EMBL:CO497151 upland cotton cDNA), EMBL:CO496924 (upland cotton cDNA), EMBL:CO496907 (upland cotton cDNA), EMBL:CO496748 (upland cotton cDNA), EMBL:CO496747 (upland cotton cDNA), EMBL:CO496373 (upland cotton cDNA), EMBL:CO496003 (upland cotton cDNA), EMBL:CO495974 (upland cotton cDNA), EMBL:CO495570 (upland cotton cDNA), EMBL:CO494691 (upland cotton cDNA), EMBL:CO493817 (upland cotton cDNA), EMBL:CO493779 (upland cotton cDNA), EMBL:CO493750 (upland cotton cDNA), EMBL:CO492925 (upland cotton cDNA), EMBL:CO492917 (upland cotton cDNA), EMBL:CO492529 (upland cotton cDNA), EMBL:CO492496 (upland cotton cDNA), EMBL:CO493902 (upland cotton cDNA), EMBL:BQ412019 tree cotton cDNA clones GA_Ed0053A09r), EMBL:CO497086 (upland cotton cDNA), EMBL:CO494597 (upland cotton cDNA), EMBL:CO494374 (upland cotton cDNA), EMBL:CO499615 (upland cotton cDNA), EMBL:CO490812 (upland cotton cDNA), EMBL:CO493178 (upland cotton cDNA).The sequence of all citations is integrated with this paper by reference.Although the none SusC albumen completely of encoding in these sequences, these different pieces can be as identifying the proteic probe of other SusC.These different pieces of this cDNA also can be used for the nucleic acid of reconstruct coding C type sucrose synthase.
Therefore the variant form that is suitable for sucrose synthase C isotype of the present invention can have following aminoacid sequence, and described aminoacid sequence comprises the aminoacid sequence that has about at least 60% or about 70% or about 80% or about 90% or about 95% sequence identity with N end consensus sequence (SEQ ID No 16) or C end consensus sequence (SEQ ID No 15) or both aminoacid sequences.Variant can have such aminoacid sequence, and the aminoacid sequence of the SusC isotype of any among described aminoacid sequence and the SEQ ID Nos.:2,3,5,7,9,11 or 13 has about at least 60% or about 70% or about 80% or about 90% or about 95% sequence identity.The nucleotide sequence of this type of variant of encoding can have such nucleotide sequence, and the nucleotide sequence of any in described nucleotide sequence and SEQ ID Nos.:1,4,6,8,10,12 or 14 the nucleotide sequence has about at least 60% or about 70% or about 80% or about 90% or about 95% sequence identity.
For the purposes of the present invention, " sequence identity " (the representing) of two associated nucleotides or aminoacid sequence with percentage ratio be meant that the number (x100) that has the position of identical residue in the sequences of two best comparisons is divided by by the number than the position.Room (that is, than being centered in the position that has residue in the sequence and do not have residue in another sequence) is considered to be the position with identical residue.Carry out the comparison of two sequences by Needleman and Wunsch algorithm (Needleman and Wunsch 1970).Can use the standard software program for example GAP (it is Wisconsin PackageVersion 10.1 (Genetics Computer Group, Madision, Wisconsin, USA) a part), use the room of default rating matrix and 50 to produce the room extension point penalty of point penalty and 3, carry out the auxiliary sequence alignment of aforementioned calculation machine easily.
Whenever be clear that define the nucleotide sequence of RNA molecule by the nucleotide sequence of mentioning corresponding dna molecular, the thymus pyrimidine in the nucleotide sequence (T) all should be substituted by uridylic (U).And what be mentioned to is that dna molecular or RNA molecule will be can be obvious from the application's context.
Also can pass through external evoked sequence variations, increase sucrose synthase C type isotype or have the functional level of the sucrose synthase of similar features.For this purpose, the method that is used for the induced sequence variation known in the art that elsewhere in this specification sheets can be described is used for for example vegetable lamb of fiber production plant, and can identify in apoplast liquid or cell walls (particularly in the growth period that secondary cell wall forms), especially in the cell walls of fiber or in the apoplast liquid of these fibers, showing higher sucrose synthase C isotype activity (for example using enzymatic determination method described herein) or showing the fiber production plant of higher sucrose synthase (especially sucrose synthase C isotype) concentration.
Produce plant for some fibre, maybe advantageously, change fiber characteristics by the functional level that reduces sucrose synthase C isotype or have a sucrose synthase of similar features.This can realize easily by the gene that imports the reticent RNA of coding.
In one embodiment, reticent RNA encoding gene can encode reticent RNA molecule or inhibitory RNA molecules change fiber characteristics thereby described reticent RNA molecule or inhibitory RNA molecules can reduce the expression of the native gene of encoding sucrose synthase C isotype.This expression decreased of the gene of encoding sucrose synthase C isotype should preferably realize by post-transcriptional silencing.Yet, be clear that, even when inhibitory RNA molecules reduced SusC genetic expression by post-transcriptional silencing, this type of RNA molecule also can be showed other functions in cell, for example instruct the dna methylation of endogenous SusC gene, finally cause this genetic expression to reduce once more.In addition, can for example use the RNAi or the dsRNA of the promoter region of the endogenous SusC gene of target, reduce endogenous SusC expression of gene by Transcriptional Silencing.
Can obtain several method in the art and be used to produce reticent RNA molecule (that is, can reduce the RNA molecule of one or one group specific expression of gene during expression), comprise so-called " justice is arranged " or " antisense " RNA technology.
Therefore, in one embodiment, the mosaic gene of coding inhibitory RNA molecules is based on so-called antisense technology.In other words, the coding region of mosaic gene comprises the nucleotide sequence of at least 20 continuous nucleotides in the complementary sequence of endogenous SusC gene nucleotide series.Can (can be as described in the elsewhere among the application by comprising dna fragmentation from least 20 Nucleotide of SusC gene, separating or identify described dna fragmentation) 3 ' the terminal formation that oppositely can express promotor with plant and participate in Transcription Termination and polyadenylation regionally effectively be connected, and makes up this type of mosaic gene.Very clear, needn't know definite nucleotide sequence or perfect kernel nucleotide sequence from this type of dna fragmentation of isolating SusC gene.
In another embodiment, the mosaic gene of coding inhibitory RNA molecules is based on the so-called technology that suppresses altogether.In other words, the coding region of mosaic gene comprises the nucleotide sequence of at least 20 continuous nucleotides in the nucleotide sequence of endogenous SusC gene of plant.Can effectively be connected by comprising 3 ' the terminal zone that forms that to express promotor with plant and participate in Transcription Termination and polyadenylation in dna fragmentation from least 20 Nucleotide of SusC gene (can as the described separation of elsewhere among the application or as described in identifying dna fragmentation) forward ground, make up this type of mosaic gene.Again, very clear, needn't know definite nucleotide sequence or perfect kernel nucleotide sequence from this type of dna fragmentation of isolating SusC gene.
Above-mentioned mosaic gene can further be strengthened by comprising following DNA element in the effect that reduces aspect the endogenous SusC genetic expression, and that described DNA element can cause is unusual, the inhibitory RNA molecules of polyadenylation is expressed or caused inhibitory RNA molecules to be trapped in the nucleus of cell.This type of DNA element that is fit to this purpose is the DNA zone of the coding of description among the WO 00/01133 (integrating with this paper by reference) from splicing ribozyme.Another this type of DNA element that is suitable for this purpose is to appraise and decide the position or be detained the DNA zone of signal with the middle coding RNA of describing of PCT/AU03/00292 (integrating with this paper by reference) that WO03/076619 announces..
A kind of convenience of downward modulation destination gene expression and very effective method are to use for example middle so-called double-stranded RNA (dsRNA) or the RNA interfering of describing (RNAi) of WO99/53050 (integrating with this paper by reference).In this technology, the RNA molecule is imported vegetable cell, wherein the RNA molecule can form and stride the about at least 19 double-stranded RNA zones to about 21 Nucleotide, wherein in two of this double-stranded RNA zone chains one list and target gene roughly the same (" the justice district is arranged ") at nucleotides sequence, and another chain lists with target gene or the complementary sequence roughly the same (" antisense district ") in justice district is arranged at nucleotides sequence.Expection is for reticent target gene expression, and the nucleotide sequence of the sequence of these 19 continuous nucleotides can have a mispairing, or has justice and antisense district can differ a Nucleotide.For realizing the structure of this type of RNA molecule or coding sex-mosaicism gene, can use the carrier of describing among the WO 02/059294.
Therefore, in one embodiment of the invention, provide to be used to change for example method of the fiber characteristics of cotton of fiber production plant, this method comprises the step that mosaic gene is imported the cell of fiber production plant, and wherein mosaic gene comprises the DNA element of following effective connection:
(a) plant can be expressed promotor, and the preferred preferentially plant that control is transcribed in fibrocyte can be expressed promotor;
The DNA zone of (b) transcribing produces the double stranded rna molecule of the endogenous SusC genetic expression that can reduce the fiber production plant when described DNA transcribes, described RNA molecule comprises the first and second RNA zones, wherein
I) nucleotide sequence that comprises with endogenous SusC gene of a RNA zone has about at least 94% nucleotide sequences sequence identity, at least 19 continuous nucleotides;
Ii) the 2nd RNA zone comprises these at least 19 the continuous nucleotide complementary nucleotide sequences with a RNA zone;
Iii) the first and second RNA zones can base pairing, thereby forms double stranded rna molecule between these at least 19 continuous nucleotides in first and second zones; With
(c) be included in the 3 ' stub area that has the Transcription Termination and the polyadenylation signal of function in the cell of plant.
The length in the first or the 2nd RNA zone (justice or antisense district are arranged) can change to the length that the equals endogenous SusC gene scope of (representing with Nucleotide) at about 19 Nucleotide (nt).The total length that justice or antisense base sequences are therefore arranged can be 25nt at least, or at least about 50nt, or at least about 100nt, or at least about 150nt, or at least about 200nt, or at least about 500nt.Expection does not have the upper limit for the total length that justice or antisense base sequences are arranged.Yet owing to putting into practice reason (for example stability of mosaic gene), expection has the length of justice or antisense base sequences should be no more than 5000nt, should be no more than 2500nt especially and can be defined in about 1000nt.
The total length that is appreciated that justice or antisense zone is long more, requires to get over not strict to the sequence identity between corresponding sequence or its complementary sequence in these zones and the endogenous SusC gene.Preferably, purpose nucleic acid should have and corresponding target sequence about at least 75%, especially about at least 80%, more particularly about at least 85%, more particularly about 90%, about especially sequence identity of 95%, more particularly about 100%, particularly corresponding section or its complementary sequence with target sequence is identical.Yet preferred purpose nucleic acid always comprises about 19 continuous nucleotides, about 25nt especially, and about 50nt more particularly, about 100nt especially, approximately the sequence of 150nt has sequence identity with the corresponding section 100% of target nucleic acid especially especially.Preferably,, the number in room is minimized, particularly for the short adopted sequence that has for sequence of calculation identity and design have justice or antisense sequences accordingly.
According to the disclosure of WO 99/53050 (integrating with this paper by reference), the mosaic gene of coding dsRNA of the present invention can comprise the intron in the intervening sequence that for example has between justice and the sense-rna zone, for example allos intron..
In another embodiment, reticent RNA or inhibitory RNA molecules can be at the microRNA molecules of target design, synthetic and/or adjusting, and it can for example cause the cutting of endogenous SusC gene the fiber production plant in the vegetable lamb.Described and produced and use several different methods (to include but not limited to people such as Schwab (2006 at the miRNA of particular target gene, Plant Cell, 18 (5): 1121-1133), WO2006/044322, WO2005/047505, EP 06009836, integrate with this paper by reference).Usually, in the target gene identification division, change existing miRNA support so that the RNA molecule that the miRNA that produces can instruct the cutting of RISC mixture to transcribe from target nucleic acid now.Can change or synthetic miRNA support, so that this miRNA comprises 21 continuous nucleotides of susC coding nucleotide sequence, the sequence shown in the sequence table for example, and allow mispairing according to described rule below this specification sheets.The specially suitable sequence that these 21 continuous nucleotides can be selected from comprises the nucleotide sequence of the SusC specific amino acid described herein of encoding..
Therefore, in one embodiment, the invention provides the expression that is used to reduce plant or increase the method for plant to the resistance of unfavorable growth conditions, the method comprising the steps of
A. mosaic gene is imported the cell of described plant, described mosaic gene comprises the DNA zone of following effective connection:
I. plant can be expressed promotor;
Ii. import vegetable cell and in vegetable cell, transcribe after be processed to the DNA zone of miRNA, wherein the miRNA mRNA that can discern the endogenous SusC encoding gene of plant and guides the cutting to this mRNA; With
Iii. randomly, participate in 3 ' DNA zone of Transcription Termination and polyadenylation.
The DNA zone of the described miRNA of being processed into can comprise following nucleotide sequence, and the nucleotide sequence of at least 21 continuous nucleotides of described nucleotide sequence and SusC encoding gene is complementary basically, and condition is to allow one or more following mispairing:
5 ' the terminal nucleotide of a.miRNA and the mispairing between the corresponding nucleotide sequence in the RNA molecule;
Mispairing in any Nucleotide in position 1 to the position 9 of b.miRNA and the RNA molecule between the corresponding nucleotide sequence;
C. three in position 12 to the position 21 of miRNA any Nucleotide and the mispairing between the corresponding nucleotide sequence in the RNA molecule, condition is not exist to surpass 2 continuous mispairing.
The functional level that reduces sucrose synthase C isotype may be favourable in the fibrocyte that for example causes linters in cotton, can reduce or avoid energy and metabolite to disperse to be used for the more disadvantageous linters of generation and to damage velveteen production.
As used herein, " native gene " is naturally occurring gene in being selected for the fiber production plant species that changes fiber characteristics, or natural existence but can import the gene that has been selected in the fiber production plant species that changes fiber characteristics by traditional breeding method in another fiber production plant species.
Also very clear, also can use mosaic gene described herein to reduce endogenous SusC expression of gene, the DNA of target-specific RNA of wherein encoding has the nucleotide sequence of at least 20 continuous nucleotides, described nucleotide sequence is selected from the nucleotide sequence of coding SEQ ID No.2,3,5,7,9,11 or 13 aminoacid sequence or their complementary sequence, or wherein target-specific RNA has the nucleotide sequence of at least 20 continuous nucleotides, and described nucleotide sequence is selected from SEQ ID No.1,4,6,8,10,12 or 14 nucleotide sequence.
As used herein, term " promotor " is meant that the RNA polymerase that is relied on DNA in the initiating process of transcribing is discerned and any DNA of combination (directly or indirectly).Promotor comprises the binding site of transcription initiation position and transcription initiation factor and RNA polymerase, and can comprise gene expression regulation albumen various other sites of bonded (for example, enhanser) thereon.
As used herein, term " plant can be expressed promotor " is meant and can controls the dna sequence dna that (initial) transcribe in vegetable cell.The promotor of mosaic gene described herein can be connected with the coding region natively, or it can be an allogeneic promoter.The term plant can be expressed any promotor that promotor comprises plant origin, and can in vegetable cell, instruct any non-plant of transcribing to come origin promoter, promptly, some promotor of virus or bacterial origin, for example CaMV35S, underground clover disease virus promoter No 4 or No 7 or T-DNA gene promoter etc.
Plant can express promotor can be composing type or the transcribing of its zone that can connect with the initial downstream of space or time mode.Preferentially the plant that the control initial sum of transcribing is kept in fibrocyte can to express promotor be such promotor, with in other cell or tissues of plant, compare the DNA regional transcription that described promotor effectively is connected with higher horizontal drive in fibrocyte and following epidermic cell.This type of promotor comprises from the promotor of the fiber-specific beta microtubule protein gene of cotton (as describing among the WO0210377), promotor (as describing among the WO0210413) from the fiber-specific actin gene of cotton, promotor (as describing among the US5792933) from the fiber-specific lipid transfer protein gene of cotton, from the promotor (WO9830698) of the expansion protein gene of cotton or from the promotor of the fiber-specific gene of describing among the promotor (US2003106097) of the chitinase gene of cotton or US6259003 or the US6166294.Promotor can also be compound (usually with transcriptional activator a unite) induction type, described in for example WO93/21334, US5514578, EP0823480, WO98/05789, WO01/34821 or WO02/20811.
Method described herein will change the feature of the fiber that is produced by the plant that changes, and comprise that intensity, length, fiber fineness, fiber maturity ratio, immature fibre content, fibre uniformity and mark swell Buddhist nun's value.
It is to depend on that the fiber maturity (or cell wall thickness of being determined by the secondary wall content of cellulose) and a kind of no unit of Fibre diameter measure (unitless measurement) that-fiber mark is swelled Buddhist nun's value (can by HVI determine).
-fibrous bundle intensity (determining by HVI) is with (cN/tex) unit representation.It is the specific tenacity of fibrous bundle, and wherein ultimate fibre fineness (tex) is worth calculating according to the grand Buddhist nun of mark.
-fiber fineness (determining by AFIS) is expressed as (mTex).What it represented a km fiber is the weight of unit with the milligram.Km fiber with 1 milligram of quality equals 1millitex.
The degree (depending on the secondary cell wall fibrin deposition) that-fiber maturity ratio (determining by AFIS) expression cell walls thickens.It is for having the ratio of the fiber of 0.5 (or bigger) circularity divided by the amount of the fiber with 0.25 (or littler) circularity.(fiber with thicker cell walls after drying more difficult flat contract and keep round).The ripening degree ratio is high more, and fiber is ripe more, and fiber is good more with regard to dyeing..
-immature fibre content (" IFC% " determines by AFIS) is the percentage ratio with the fiber that is lower than 0.25 ripening degree.IFC% is low more, and fiber is applicable to dyeing more.
-several different units are used as the index of staple length, show existing wherein three the value of describing.
Upper half mean length (upper half mean) (" UHM ", by HVI determine) is the mean length (weight bias) of the longest half fiber.
-fibre uniformity index (" UI " determines by HVI) expression mean value (mean length) is to the ratio of upper half mean length.It is measured for staple length dispersive in the colony; If all fibres length is identical, then UI will equal 100%.Short fibre content (Short Fiber Content) (" SFC% " passes through HVI) is based on the weight meter and is shorter in length than [1/2] " the percentage ratio of fiber.HVI is considered to measure the short fibre content of only being determined by genetics, and this is because this measurement does not apply extra potential fibre breakage stress.
Other staple length indexs comprise the length (weight basis length) (" L (w) " [in] determines by AFIS) based on weight, and it is based on the mean length of the fiber of weight calculating.Length (number basis length) (" L (n) " [in] determines by AFIS) based on number is the mean length by the fiber of number calculating.Length " L5% (n) " [in] (determining by AFIS) is 5% the length of span, or when fiber the length of 5% fiber leap during parallel and stochastic distribution.Length " L2.5% (n) " [in] (determining by AFIS) is 2.5% the length of span, or when fiber the length of 2.5% fiber leap during parallel and stochastic distribution." UQL (w) " [in] (determining by AFIS) is by weight the upper quartile length of the fiber of calculation, or calculates by weight the length that 25% fiber surpasses." SFC (n) " [in] and " SFC (w) " [in] at last (determining by AFIS) is respectively the percentage ratio that is shorter than 0.50 inch long fiber on number and weight basis.Opposite with HVI, AFIS can hit fiber before adopting these measurements, and this may cause fibre breakage.Therefore, AFIS SFC value is the good index of fiber characteristics after the normal process.
Very clear, can be with for example ways and means of the fiber characteristics in the vegetable lamb and the additive method combination of change fiber characteristics known in the art of change fiber production plant described herein.
In one embodiment of the invention, with the ways and means combination of describing among the application's ways and means and WO2005/017157 (integrating with this paper by reference) or the WO01/17333.Expect that the combination expression of these genes will cause synergistic effect in the increase of staple length and/or intensity.Also can be with additive method and the means combination of describing among the application's ways and means and for example PCT/EP2006/005853 or the WO98/00549 that fiber characteristics changes that relate to.
These mosaic genes can maybe can be integrated with mosaic gene in the cell of a plant by the hybridization between the plant of each self-contained mosaic gene by transforming the cell that imports a plant continuously.
Because the fiber for example secondary cell wall of cotton fibre is synthetic initial overlapping with the fiber extended peroid, thus may be further advantageously, it is initial until than late period to postpone the expression of sucrose C isotype, thus prolongation fiber extended peroid.Also this delay of SusC genetic expression can be expressed in the increase in this late period with the SusC gene and be combined.A method that obtains this delay is for opening the SusC gene of expressing in later period than this endogenous SusC gene with endogenous SusC gene " transposing ", for example, by as described herein, reduce endogenous SusC genetic expression, and introduce and to be the promotor that the later etap of secondary cell wall synthetic opens (E6 promotor (John ME for example, Keller G, Proc Natl Acad Sci USA1996,93:12768-12773)) the SusC coding region under the control.Can dystopy import the chimeric susC coding region that be under the control of secondary cell wall expression promoter in synthetic late period, maybe can import endogenous SusC gene by the homologous recombination technique of describing among the PCT/EP2006/003086 for example.Owing to compare with the susC gene of upland cotton Cultivar, the sea island cotton Cultivar for example susC gene of PIMA kind shows in later expression in period, can prolong the fiber extended peroid and cause longer cotton fibre so expection is used from the susC gene in susC gene " transposing " the upland cotton Cultivar of sea island cotton Cultivar.
The present invention also comprises mosaic gene described herein, and comprises plant, seed, the tissue of these mosaic genes and the fiber that produces from this type of plant.
The method that transforms plant is known in this area.The method of converting cotton plant is also known in this area.In for example United States Patent (USP) 5,004,863 or United States Patent (USP) 6,483,013, described the agriculture bacillus mediated conversion of cotton, in WO 92/15675, reported the cotton that is undertaken by microparticle bombardment and transformed.
Can use the method for knowing in this area, mosaic gene of the present invention be imported plant with stable manner or in instantaneous mode.Mosaic gene can be imported plant, or can described in EP 1339859, in vegetable cell, produce mosaic gene.
Can mosaic gene be imported the cotton plant that can produce embryo (embryogenic) callus by transforming, Coker312 for example, Coker310, Coker 5Acala SJ-5, GSC25110, FIBERMAX819, Siokra 1-3, T25, GSA75, Acala SJ2, Acala SJ4, AcalaSJ5, Acala SJ-C1, Acala B1644, Acala B1654-26, Acala B1654-43, AcalaB3991, Acala GC356, Acala GC510, Acala GAM1, Acala C1, AcalaRoyale, Acala Maxxa, Acala Prema, Acala B638, Acala B1810, AcalaB2724, Acala B4894, Acala B5002, non Acala " picker " Siokra, " stripper " kind FC2017, Coker315, STONEVILLE506, STONEVILLE825, DP50, DP61, DP90, DP77, DES119, McN235, HBX87, HBX191, HBX107, FC3027, CHEMBRED A1, CHEMBRED A2, CHEMBRED A3, CHEMBRED A4, CHEMBRED B1, CHEMBRED B2, CHEMBRED B3, CHEMBRED C1, CHEMBRED C2, CHEMBRED C3, CHEMBRED C4, PAYMASTER 145, HS26, HS46, SICALA, PIMA S6ORO BLANCOPIMA, FIBERMAX FM5013, FIBERMAX FM5015, FIBERMAXFM5017, FIBERMAX FM989, FIBERMAX FM832, FIBERMAX FM966, FIBERMAX FM958, FIBERMAX FM989, FIBERMAX FM958, FIBERMAX FM832, FIBERMAX FM991, FIBERMAX FM819, FIBERMAX FM800, FIBERMAX FM960, FIBERMAX FM966, FIBERMAX FM981, FIBERMAX FM5035, FIBERMAX FM5044, FIBERMAX FM5045, FIBERMAX FM5013, FIBERMAX FM5015, FIBERMAX FM5017 or FIBERMAX FM5024, and have the genotypic plant that derives from it.
Herein, " cotton " comprises upland cotton or sea island cotton." cotton ancestors plant " comprises the cotton and long calyx cotton of tree cotton, cotton and Lei Mengdeshi.
Yet, ways and means of the present invention also can be used for other plant species for example hemp, jute, flax and xylophyta, includes but not limited to Pinus (Pinus spp.), Populus (Populus spp.), Picea (Picea spp.), eucalyptus genus (Eucalyptus spp.) etc.
The plant transformed that obtains can be used for conventional breeding system producing the conversion plant that more has same characteristic features or mosaic gene of the present invention is imported other kinds of identical or corresponding plants species, or imports hybrid plant.The seed that obtains from plant transformed comprises mosaic gene of the present invention with the form of stable genome inset, and is also included within the scope of the present invention.
In another embodiment, the colony that provides of being used at specified plant species (fiber production plant species) to have different genotype or kind identifies the method for the proteinic allelic variation that relates to fiber characteristics, wherein said protein or quantity and/or the quality with fiber production is relevant individually or in combination.This method comprises the following steps:
A) provide one of the specified plant species to have different varieties or genotypic colony, perhaps (for example at the sucrose synthase C isotype coding nucleotide sequence that comprises different allelic forms, coding SEQ ID NO 1,4,6,8,10,12 or 14 nucleotide sequence) species hybriding between the plant species.Can use the method that elsewhere is described among the application to identify different allelic forms.Preferably, provide segregating population, wherein have the various combination of the proteic allelic variation of SusC.The method that produces segregating population is known in field of plant breeding;
B) determine each individual fiber characteristics correlation parameter of colony;
C) determine the existence of the specific allelic form of the nucleotide sequence of coding SusC in each individuality of colony; With
D) existence with the particular combinations of the existence of the specific allelic form of the appearance of special fiber feature and described nucleotide sequence or this type of allelic form associates.
Can use the information of gained, determine the existence of allelic form or do not exist, quicken to have the procedure of breeding of the kind of special fiber or drought resisting feature by using conventional Protocols in Molecular Biology.
The allelic form of the SusC gene that can also identify, separation is relevant with the special fiber feature also imports for example vegetable lamb of plant with it, and wherein endogenous SusC expression of gene is reduced or eliminates.This minimizing of endogenous SusC genetic expression can realize by post-transcriptional silencing described herein or Transcriptional Silencing easily, maybe can for example lack endogenous SusC gene by inactivation and realize.Can import allelic form by breeding technique or by using isolating gene transformation.
In another embodiment of the invention, the antibody that provides anti-this novel type of sucrose synthase gene to produce, particularly identification have the proteic antibody of SusC of the aminoacid sequence of SEQ ID No.s 2,3,5,7,9,11 or 13.
" comprise " feature, integer, step or the composition that should be construed as denoting existence and address as used herein, but do not get rid of existence or add one or more features, integer, step or composition or their group.Therefore, for example, the nucleic acid or the protein that comprise Nucleotide or aminoacid sequence can comprise more Nucleotide or the amino acid of quoting from than reality, that is, be included in the bigger nucleic acid or protein.The mosaic gene that comprises the definite DNA zone of structure or function can comprise extra DNA zone etc.
Following indefiniteness embodiment has described the evaluation that participates in secondary plant cell wall synthetic novel type of sucrose synthase and has been used to change for example mosaic gene and uses thereof of the fiber characteristics of cotton of fiber production plant.Unless point out in addition in an embodiment, otherwise according to people such as Sambrook (1989) Molecular Cloning:A Laboratory Manual, the 2nd edition, Cold Spring HarborLaboratory Press, people such as NY and Ausubel (1994) Current Protocols inMolecular Biology, Current Protocols, the standard scheme of describing in the 1st and 2 volumes of USA carries out all recombinant DNA technologies.By BIOS Scientific Publications Ltd (UK) and Blackwell Scientific Publications, among the Plant MolecularBiology Labfax (1993) of UK combined publication (R.D.D.Croy work) standard material and the method that is used for the plant molecular operation described.
In whole specification sheets and embodiment, mention the following sequence shown in the sequence table:
SEQ ID No.:1: coding is from the nucleotide sequence of the cDNA of the sucrose synthase C type of cotton.
SEQ ID No.:2: from the aminoacid sequence of the sucrose synthase C type of the cDNA of cotton coding.
SEQ ID No.:3: from the aminoacid sequence of the SusC of upland cotton cultivar.
SEQ ID No.:4: from the nucleotide sequence of the SusC genomic dna of upland cotton cultivar.
SEQ ID No.:5: from the aminoacid sequence of the SusC of sea island cotton cultivar.
SEQ ID No.:6: from the nucleotide sequence of the SusC genomic dna of sea island cotton cultivar, the SUS C albumen of coding SEQ ID No.:7.
SEQ ID No.:7: from the aminoacid sequence of the SusC of tree cotton.
SEQ ID No.:8: from the nucleotide sequence of the SusC genomic dna of tree cotton.
SEQ ID No.:9: from the aminoacid sequence of the SusC of Lei Mengdeshi cotton.
SEQ ID No.:10: from the nucleotide sequence of the SusC genomic dna of Lei Mengdeshi cotton..
SEQ ID No.:11: from the aminoacid sequence of the SusC of the cotton hypotype 1 of long calyx.
SEQ ID No.:12: from the nucleotide sequence of the SusC genomic dna of the cotton hypotype 1 of long calyx
SEQ ID No.:13: from the aminoacid sequence of the SusC of the cotton hypotype 2 of long calyx
SEQ ID No.:14: from the nucleotide sequence of the SusC genomic dna of the cotton hypotype 2 of long calyx
The aminoacid sequence of the C end consensus sequence of SEQ ID No.:15:SusC
The aminoacid sequence of the N end consensus sequence of SEQ ID No.:16:SusC
SEQ ID No.:17: as the oligonucleotide sequence (5 ' UTR) of SUS C Auele Specific Primer
SEQ ID No.:18: as the oligonucleotide sequence (3 ' UTR) of SUS C Auele Specific Primer
Embodiment
The evaluation of C type sucrose synthase in the embodiment 1. Gossypium species
The evaluation of SusC.The alleged occurrence at least 3 classes Sus gene of expressing between the growth period in cotton fibre is called A, B and C type.A/D-type and sequence (accession number U73588) homology of reporting before, Type B show and the sequence of this announcement has strong sequence similarity.The C type sequence that provides in this specification sheets is new sequence and shows differential expression at extended peroid and secondary cell wall between the phase.It is the isotype than the appearance in late period of growing at fiber that the C type shows as.SusC has only 76% identity with A and Type B albumen on amino acid levels.By using RTPCR and Northern to analyze, the growth express spectra that has confirmed C type transcript but demonstrated and the pattern that more matches in this fiber effect in the growth period before secondary cell wall is synthetic, and A and Type B are expressed at the early stage height that fiber extends.C type gene is strong expression in the stem tissue also.
Different steps in the fiber growth: cDNA is separated from fibrous tissue in the back 5,10,15,20,40 days (dpa) of blooming.Produce the library from these cDNA, and DNA isolation.Then, use this DNA, use the parent material of 9ng, characterize susyC and express as PCR reaction (this reaction is used the SusC type specificity primer of SEQ ID Nos:17 and 18 as primer) with each library DNA.The expection size of amplified fragments is 2550bp for cDNA, is 3353bp for genomic dna.Carry out suitable contrast to verify the sxemiquantitative feature of this assay method.Fig. 2 display result, it is abundanter relatively at 20-40dpa to draw SusC mRNA by it.
Protein fractionation separates.Fractional separation is from the albumen of the fiber extraction of extended peroid (8-11DAF) and secondary cell wall synthesis phase (15-25DAF), so that solubility (kytoplasm) and microsome (rich plasma membrane, cytoskeleton and vacuole skin) fraction to be provided.Be further purified microsomal fraction with the enrichment plasmalemma protein.Judge that as western trace this PM fraction no longer has any cytoskeleton or cell walls pollutes by anti-Actin muscle.Also comprise another cell fraction, described cell grade branch comprises the apoplast albumen that obtains from complete fiber by with the gentle washing of the isotonic buffer solution that contains proteinase inhibitor and EGTA.These protein Preparation things are carried out electrophoresis on SDS-PAGE, identify Sus albumen by the western trace.Cut gel fragment, it is carried out tryptic digestion, identify the form of the Sus that exists then by tandem mass spectrum.The example that is used to identify the sign peptide of SusC isotype is shown in table 1A and 1B.Different with the proteic 92.4kDa of Type B with A/D, the predicted molecular weight of Sus-C is 90.3kDa, thereby allows to differentiate on SDS-PAGE C type and A/B isotype.Can analyze these 2 by peptide MS-MS and bring this resolution of affirmation (bigger band comprises the sign peptide from A and B, and less band has unique C type peptide individually).
Table 1A: detected SusC specific peptide in apoplast
The 8th day apoplast The 17th day apoplast The 25th day apoplast
Do not detect the C type 9.20e+005 (R)KAEEYLTPLSSDTPYSVFEK() 1.45e+006 (R)IQDVNSLQHALR()
8.17e+005 (K)LLTLTGVYGFSK(H) 2.37e006 (R)KAEEYLTPLS SDTPYSVFEK()
1.32e+006 (K)YTWQIYSEK() 2.16e006 (K)NLTGLVEFYAK(N)
6.90e+005 (R)STQEAVVSSPLVALAIR()
7.45e+006 (K)LQGLDITPR(I)
3.64e+006 (K)YTWQIYSEK()
5.53e+006 (R)VVDGIDVFDPK(F)
3.34e+006 (K)LLTLTGVYGFSK(H)
4.17e+005 (K)D TLGQYESHIAFTLPGLYR(V)
3.50e+005 (K)LFVEEMPVAEYLR(L)
Table 1B: detected SusC specific peptide in microsomal fraction
The 8th day microsome The 13rd day microsome The 20th day microsome The 25th day microsome
Do not detect the C type Do not detect the C type 6.06E+006 (R)KAEEYLTPLSSDTPYSVF EK() 3.94E+006 (R)KAEEYLTPLS SDTPYSV FEK()
7.64E+006 (K)LLTLTGVYGF SK(H) 1.13E+006 (K)LFVEE MPVAEYLR(L)
2.31E+006 (R)IQ DVNSLQHALR() 3.89E+006 (R)VVDGIDVFDPK(F)
6.70E+006 (R)VVDGIDVFDPK(F 3.68E+006 (K)LLTLTGVYGF SK(H)
1.82E+006 (R)YIEmLHAWmYNNR(V) 3.97E+005 (K)SGFNIDPYNGDLAAETLANF FEK(C)
3.19E+006 (K)YTWQIYSEK() 2.63E+006 (K)NLTGLV EFYAK(N)
1.99E+006 (R)STQEAVVSSPLVALAIR() 1.26E+006 (R)STQEAVV SSPLVALAIR(
1.05E+006 (R)LGESLATHPQQAK(S) 2.05E+006 (K)NLTGLVEFYAK(N)
5.72E+006 (K)LFVEE mPVAEYLR(L) 3.01E+006 (K)D TLGQYESHIA FTLPGLYR(V)
8.25E+005 (K)DTLGQYESHIAFTLPGLY R(V) 2.18E+006 (R)IQ DVNSLQHALR()
5.25E+006 (K)LQGLDITPR(I) 8.02E+005 (R)YIEmLH AWmYNNR(V)
3.28E+006 (K)NLrGLVEFYAK(N) 3.60E+005 (K)TKYPDSDIN WK(Q
9.91E+005 (K)SIGNGmDFLNR(H)
5.35E+005 (R)L GESLATHPQQ AK(S)
5.59E+005 (R)ALEEELLHR(F)
2.77E+006 (K)LFVEE mPVAEYLR(L)
Table 2 has been summed up the data of mass spectrum and SDSPAGE/Western.It should be noted that proteic low abundance especially from the Sus-C of solubility, microsome and PM fraction.Consider at fiber and grow that this is seemingly irrational than there being high-caliber transcript late period.Growing than carrying out the apoplast washing late period, showing that C type albumen is present on the western trace with high level (with respect to A), this is with opposite for the observations of kytoplasm, microsome and PM fraction.Sus-C is highly abundant at 25DAF, and can not detect by peptide MS-MS at 8DAF.On the contrary, Sus-A and possibly Sus-B between the growth period be ubiquitous (in all fractions).Sus-A is main in the PM fraction, and unique possibly isotype, is shown as the protein of 85kDa on SDS-PAGE.
Table 2: the summary of mass spectrum and SDSPAGE/Western data
8dpa microsome (top band) Susy A+B
8dpa microsome (following band) Susy A or A+B
The 15dpa microsome Susy A+C or A+B+C
The 8dpa plasma membrane Susy A or A+B
The 15dpa plasma membrane SusyA or A+B
The 8dpa solubility Susy A or A+B
The 15dpa solubility Susy A+B
8dpa apoplast (top band) Susy A+B (low-down amount)
8dpa apoplast (following band) Susy A+B (main band)
25dpa apoplast (top band) Susy A+B+C ＊(low-down amount)
25dpa apoplast (following band) Susy C (main band)
*The existence of C in this sample is probably owing to the pollution from the big band below SusC.
Sus-C in the apoplast fraction of the number of peptide and 25DAF is complementary, and the western signal shows that this isotype is very abundant, and only mainly is present in the apoplast of 25DAF.Enjoyably, SusA and B are present in the apoplast of 8DAF fiber as the advantage form.These results hint that Sus plays a major role in the cell walls of cotton fibre, and the expression of Sus-C on rna level is relevant with the formation of secondary cell wall, and this proteic abundance prompting Sus-C is the main cell walls isotype of secondary cell wall synthesis phase in the cell walls of 25DAF.EM immunity metal working clearly illustrates the signal at the Sus of secondary cell wall outside.This is with in full accord in secondary cell wall stage Sus-C albumen existing in apoplast.
Embodiment 2. analyzes the SusC isotype and identifies that cotton is different with the sucrose synthase in the relevant species Difference between the isotype.
Phosphorylation site: at length characterized a serine phosphorylation site that is positioned at this proteinic N-terminal in the literature, it has the consensus sequence of RXXS.Corn (Zea mays) (have a following site: 12RXXS 15) in this specific Serine phosphorylation as if with the susy enzyme from the release of plasma membrane relevant (people such as Winter H, 1997-FEBS Letters 420,151-155; People such as Hardin SC, 2004, Plant Physiology 134,1427-1438).In the site 8RXXS 11The sudden change of going up this phase homoserine cause the mung bean sucrose synthase use sucrose to produce to have more aspect the UDP glucose effect (=higher Km) (people such as Nakai T., 1999, Proc.Natl.Acad.Sci.USA 96,14-18).Also characterized the phosphorylation on this specific site in the soybean, as if described phosphorylation relevant with hydrophobic forfeiture, infer thus with plasma membrane combine attenuation of correlation (people such as Zhang X.Q., 1999 Archives ofBiochemistry and Biophysics 371,70-82).Also at the other plant species: plane (sycamore) (people-2004-Physologia Plantarum 122 such as Pozueta-Romero J., p.275), Japanese pear (Japanese pear) (people-2002-PhysologiaPlantarum 114 such as Tanase K., p.21) and upland cotton (upland cotton) (people-2001-Plant Molecular Biology 47 such as Haigler C.H. has characterized this phosphorylation site in 29-51).The conclusion that draws from these researchs shows that phosphorylation has influence to the Km value of enzyme.
All can identify similar consensus sequence in the separative sucrose synthase C albumen with following amino acid sites: 27RXXS 30Therefore, this phosphorylation site is positioned at proteinic more the inside.Can be at other known plant sucrose synthase genes: find similar sign among L19762, AY205302, AY205085, AJ537575, M18745, U2487, X75332, Y76091, X82504, the AJ131999.These genes most of have this two phosphorylation sites simultaneously: 8RXXS 11With 27RXXS 30Protein L19762 and AJ131999 only have the latter.
Characterized another serine phosphorylation site in the document: from the Ser170 of sus1 (corn).This site and label susy-albumen with carry out proteolysis relevant (people such as Hardin S.C., 2003, The Plant journal 35,588-603).This Ser also is present among the SusC.Have only two sucrose synthases not comprise this serine residue: Atsus5 and AB018561.
Be present in protein domain, functional site and specific primitives (using Inter ProScan program) among the SusC.SusC comprises two structural domain and the family zones that are called sucrose synthase (amino acid/11 8 to 794) that are called sucrose synthase (from amino acid/11 to 545) and glycosyltransferase (from amino acid 549 to 737).
The UDP-binding site: following sequence is the uridine land on the sucrose synthase C albumen: 270VVIMTPHGYFAQDNVLGYPDTGGQVVYILDQVRALEEELLHRFKLQGLDITPRILV ITRL 329This sequence shows very high homology between all known plants sucrose synthases.
Genome structure.On the genomic level of susyC gene, carry out comparison (Fig. 3) with other isolating cotton susy genes.Very clear immediately, the susyC gene has special genes group structure, because compare with other susy genes from Gossypium, it has intron still less.The intron exons structure of SusyC gene is also significantly different with Arabidopis thaliana susy gene.
Hydrophobicity.By susyC and other known plant sucrose synthases are compared, analyzed the hydrophobicity spectrum of susyC.N-terminal shows as hydrophobic, and this is an all undiscovered special feature in most of plant sucrose synthases.Following sucrose synthase is the most similar to SusC on this level: SUS6 (Arabidopis thaliana), from the sucrose synthase (D10266) of mung bean, from the sucrose synthase (DD014303) of eucalyptus with from the sucrose synthase (AY341026) of willow.
The Actin muscle land.As if described the Actin muscle land of inferring in the literature, this land is present in the sucrose synthase (people-1998-FEBS letters 430 such as Winter H., 205-208; Winter H. and Huber S.C.-2000-Critical Reviews in Plant Sciences 19 (1), 31-67).This regiospecificity is in conjunction with the F Actin muscle.The susyC Actin muscle binding site of inferring has following amino acid sequences: 383KDVAAEITKEFQ 394The amino acid outstanding with runic is the amino acid that is specific to SusyC.Consensus sequence in this site between the different plant sucrose synthases is: [K *, E, D]-D-[V, A]-[A, G, S, T]-X-E-[L, V, I]-[T, S, A, M ,]-[K, R, G, M, L]-E-[F *, L, M]-[Q, N].
Enjoyably, 383K (=lys) have a basic side chain, yet alternative amino-acid residue (E (=glu) and D (=asp)) have acid side-chain.Because K is that susyC is unique, so this may illustrate this specific protein debond F Actin muscle fibril.In addition for susyC be unique F (=Phe) amino acid recently its homologue L and the M volume from the other plant sucrose synthase is bigger.This may further stop and the combining of Actin muscle.
SusC albumen special aminoacid sequence.After different plant sucrose synthase comparisons, very clear, some zones of susyC gene are very different and special.These zones lay respectively at the N of susyC gene and C-terminal and have following amino acid sequences:
A (N end regions)= 36HKSQKLLSVLDKEAGNQALDGMVV 59
B (C end regions)= 765AYQEQRGRKRYIEMLHAWMYNNRVKT 790
By observing the 2D structure of prediction, analyzed the N end regions.For this purpose, use follow procedure: HCA (hydrophobic cluster analysis) and JPRED.To compare from the result of two programs, and compare (Fig. 4) with other sucrose synthases from upland cotton.Very clear according to this analysis, susC has very particular structure on this position.Different phosphorylation sites are also pointed out, confirm the peculiar property of susyC gene.The C end regions has been carried out similarity analysis (Fig. 5).Once more, in this zone, the structure of SusC also is different with structure from other sucrose synthases of cotton.
Use N end and the C terminal specific zone of SusC in database, to identify est sequence.The results are summarized in the following table 3.
Ovule (3 to 0) Primary cell wall (0 to 15dpa) Secondary cell wall (20 to 40dpa) Other Gossypium species The other plant species Add up to
The N end regions 0 0 35 1(＝7-10dpa) 0 36
The C end regions 1 0 30 1(＝7-10dpa) 0 32
Table: the number of the EST that extracts from some particular organizations and plant species.
Crossing of SusC expressed in embodiment 3. cottons.
By using the standard recombinant dna technology, effectively connect following DNA element:
The cotton fiber specific promoter of describing among the WO2004/066571
DNA zone encoding sucrose synthase C, that have the nucleotide sequence of SEQ ID No.1
3 ' no terminator zone.
Select gene to import the T-DNA carrier with bar this mosaic gene.The T-DNA carrier is imported agrobacterium tumefaciens (Agrobacterium tumefaciens),, be used to produce the transgenic cotton flowering plant described in 013 as US 6,483.
Analysis comprises the transgenic cotton flowering plant of mosaic gene, especially fiber, the expression of the increase of middle SusC, and, analyze the fiber that obtains from these plants with regard to the grand Buddhist nun's value of fibre strength, staple length, fiber maturity ratio, immature fibre content, fibre uniformity and mark..
The expression of downward modulation SusC isotype in embodiment 4. cottons.
By using the standard recombinant dna technology, effectively connect following DNA element:
CaMV 35S promoter zone
Corresponding to the C end SusC specific sequence of the N end SusC specific sequence nucleotide sequence of SEQ ID No 1, coding SEQ ID No 18 or SEQ ID NO 17 adopted RNA coding region arranged.
Sense-rna coding region corresponding to the complementary sequence of the nucleotide sequence that adopted RNA district is arranged.
3 ' no terminator zone..
Select gene to import the T-DNA carrier with bar this mosaic gene.The T-DNA carrier is imported agrobacterium tumefaciens,, be used to produce the transgenic cotton flowering plant described in 013 as US 6,483.
Analysis comprises the transgenic cotton flowering plant of mosaic gene, especially fiber, the expression of the minimizing of middle SusC, and, analyze the fiber that obtains from these plants with regard to the grand Buddhist nun's value of fibre strength, staple length, fiber maturity ratio, immature fibre content, fibre uniformity and mark..
Embodiment 5.SusC isotype crossing in transgenic arabidopsis expressed.
By using the standard recombinant dna technology, effectively connect following DNA element:
The CaMV35S promoter region;
The DNA zone of the SusC of coding SEQ ID No 2 (SEQ ID No 1 from nt 84 to nt 2474);
3 ' no terminator zone.
In addition, effectively connect following DNA element
Can in a series of plant species, cause tieing up the clover S2A gene of the preferential expression of pipe and trichome the stem specificity promoter (people such as Abrahams, 1995, Plant.Mol.Biol.27:413-528).
The DNA zone of the SusC of coding SEQ ID No 2 (SEQ ID No 1 from nt 84 to nt2474).
3 ' no terminator.
Select gene to import the T-DNA carrier with bar these mosaic genes.The T-DNA carrier has been imported agrobacterium tumefaciens, and by using flower-dipping method (floral dip) to be used to produce transgenic arabidopsis.The plant that regenerated, the several significant phenotypes of normal observation have write down the phenotype in T2 generation in available from the transgenic plant of one of above-mentioned transgenosis.
Many transgenic strains of two kinds of constructs all show the root branch that increases in cultivation, comprise a large amount of lateral root growths.In addition, the forfeiture of apical dominance, the symmetry of spending more bolting (multiple floralbolt), rosette-stape leafage and the destruction of compactness forfeiture, phyllotaxy and inflorescence and the flowering time of delay have been observed.
The branch that increases and the forfeiture of apical dominance are considered to extend owing to the precocious secondary cell that reduces in the meristem zone that causes that thickens owing to cell walls.
The transgenic strain with arbitrary mosaic gene of vast scale demonstrates the phenotype influence to the trichome structure.As if the branch and the thick pulvinus that increase are general visible (Fig. 6 C and D), and trichome density increases.
The stem of Sus-C transgenic strain and flower a kind of sedge demonstrate end usually and increase slightly (extremethickening), and bifurcated, fasciation (Fig. 6 A and B) possibly.
Used three kinds of methods that phenotype and transgene expression are associated:
1) Sus-C and endogenous Sus Gene RT-PCR;
2) the western trace (referring to Fig. 7) that uses the Sus-C specific antibody to carry out;
3) the active mensuration of Sus.
Show transgenic strain and the heaviest phenotypic correlation that high-caliber 35S-Sus-C expresses.
Sequence table
＜110〉Commonwealth Scientific and Industrial Research Organization
Bayer Biochemical Sciences S.A.
＜120〉evaluation of novel type of sucrose synthase and the purposes in fibre modification thereof
<130>BCS 06-2008WO1
<150>EP 06015433.3
<151>2006-07-25
<150>US 60/834066
<151>2006-07-28
<160>18
<170>PatentIn version 3.3
<210>1
<211>2635
<212>DNA
＜213〉upland cotton (Gossypium hirsutum)
<220>
<221>CDS
<222>(84)..(2474)
<400>1
acgcggggtg tctttgattt ttatccatat cctaattctt catacattcc ttcccttctg 60
ccatttcagg aacccaaaaa aca atg gct tca atc agt gtt tgt gag cgt ttg 113
Met Ala Ser Ile Ser Val Cys Glu Arg Leu
1 5 10
ggt gaa tct cta gct act cat cca cag cag gca aag tct atc ttg tca 161
Gly Glu Ser Leu Ala Thr His Pro Gln Gln Ala Lys Ser Ile Leu Ser
15 20 25
agg att gaa agc ctc gga aag ggt att cat aag tct caa aag ctg ctc 209
Arg Ile Glu Ser Leu Gly Lys Gly Ile His Lys Ser Gln Lys Leu Leu
30 35 40
tcg gtt ctc gat aaa gag gcc gga aat caa gca ctt gat ggg atg gtg 257
Ser Val Leu Asp Lys Glu Ala Gly Asn Gln Ala Leu Asp Gly Met Val
45 50 55
gtg ggg gtc ctc agg tcc act cag gaa gct gta gtg tcg tct cca ttg 305
Val Gly Val Leu Arg Ser Thr Gln Glu Ala Val Val Ser Ser Pro Leu
60 65 70
gtt gcc ctt gcc att cgt tca gct cct gga gtt tgg gag tac att gct 353
Val Ala Leu Ala Ile Arg Ser Ala Pro Gly Val Trp Glu Tyr Ile Ala
75 80 85 90
gtg gag gtc caa aag ctt ttt gtg gag gaa atg ccc gtt gct gag tat 401
Val Glu Val Gln Lys Leu Phe Val Glu Glu Met Pro Val Ala Glu Tyr
95 100 105
cta cgg ttg aag gaa gaa ctt gtt gat gga agc tcc aat ggc gag ttt 449
Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser Ser Asn Gly Glu Phe
110 115 120
atg ttg gaa ttg gac ttt ggt gca ttc aat aat tct gtt cct cgt cca 497
Met Leu Glu Leu Asp Phe Gly Ala Phe Asn Asn Ser Val Pro Arg Pro
125 130 135
tct ctt tca aag tcc att ggt aat ggc atg gac ttc ctc aac cgc cac 545
Ser Leu Ser Lys Ser Ile Gly Asn Gly Met Asp Phe Leu Asn Arg His
140 145 150
ctt tct gcc aag cta ttt caa gac aag gag aac ttg aac ttg ttg ctt 593
Leu Ser Ala Lys Leu Phe Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu
155 160 165 170
gaa ttt ctc caa att cac tgc cag aag gga aag ggt atg ctg ttg aat 641
Glu Phe Leu Gln Ile His Cys Gln Lys Gly Lys Gly Met Leu Leu Asn
175 180 185
gac aga atc caa gat gtg aat tcc ctc caa cat gca tta agg aag gcc 689
Asp Arg Ile Gln Asp Val Asn Ser Leu Gln His Ala Leu Arg Lys Ala
190 195 200
gag gag tat ctg act cct cta tcc tcg gat acc ccg tac tca gtt ttc 737
Glu Glu Tyr Leu Thr Pro Leu Ser Ser Asp Thr Pro Tyr Ser Val Phe
205 210 215
gag aaa agg ttt ctg ggg att ggt ttg gag aag gga tgg ggt gat aat 785
Glu Lys Arg Phe Leu Gly Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn
220 225 230
gct gag cat gtt ctt gag atg atc cat ctt cta ttg gat ctc ctt cag 833
Ala Glu His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu Leu Gln
235 240 245 250
gca cct gat cct gtc gca ctt gaa agc ttc ctt gga aga atc cca ttg 881
Ala Pro Asp Pro Val Ala Leu Glu Ser Phe Leu Gly Arg Ile Pro Leu
255 260 265
gtc gcc aat gtt gta atc atg act ccc cat gga tac ttt gcc caa gac 929
Val Ala Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Ala Gln Asp
270 275 280
aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat atc tta 977
Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr Ile Leu
285 290 295
gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt ttc aag ctg 1025
Asp Gln Val Arg Ala Leu Glu Glu Glu Leu Leu His Arg Phe Lys Leu
300 305 310
caa gga ctc gac att acc cca cgt atc cta gtc att act cgg ctc ctc 1073
Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val Ile Thr Arg Leu Leu
315 320 325 330
cct gat gct gta gga aca act tgc ggt cag cgt ctt gag aaa gtg tat 1121
Pro Asp Ala Val Gly Thr Thr Cys Gly Gln Arg Leu Glu Lys Val Tyr
335 340 345
gga acc aag tat tct gat att ctt cgg gta ccc ttc aga act gag aag 1169
Gly Thr Lys Tyr Ser Asp Ile Leu Arg Val Pro Phe Arg Thr Glu Lys
350 355 360
gga att gta cgt cca tgg atc tca cga ttc aaa gtc tgg cct tac ctg 1217
Gly Ile Val Arg Pro Trp Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu
365 370 375
gag act tac act aag gat gtt gca gct gag atc acc aaa gag ttc cag 1265
Glu Thr Tyr Thr Lys Asp Val Ala Ala Glu Ile Thr Lys Glu Phe Gln
380 385 390
ggc aag cct gat ctg att gtt ggg aac tac agt gat gga aac att gtt 1313
Gly Lys Pro Asp Leu Ile Val Gly Asn Tyr Ser Asp Gly Asn Ile Val
395 400 405 410
gct tct tta ttg gca cat aag ttt gat gtt aca cag tgc act att gct 1361
Ala Ser Leu Leu Ala His Lys Phe Asp Val Thr Gln Cys Thr Ile Ala
415 420 425
cgt gca ctc gag aag acg aaa tac ccg gat tca gac att aac tgg aaa 1409
Arg Ala Leu Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys
430 435 440
cag ctt gag gat aag tat cac ttc tcc tgt cag ttt act gct gat ctt 1457
Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Ala Asp Leu
445 450 455
att gct atg aac cat act gat ttt atc atc acc agc acc ttc caa gag 1505
Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu
460 465 470
att gct gga agc aag gac act ctc ggc caa tac gag agt cac att gct 1553
Ile Ala Gly Ser Lys Asp Thr Leu Gly Gln Tyr Glu Ser His Ile Ala
475 480 485 490
ttc act ctt cca ggg ctc tac cgc gtt gtt gat ggg atc gat gtt ttt 1601
Phe Thr Leu Pro Gly Leu Tyr Arg Val Val Asp Gly Ile Asp Val Phe
495 500 505
gat ccc aaa ttc aat att gtc tcc cct ggt gct gat atg agc ata tac 1649
Asp Pro Lys Phe Asn Ile Val Ser Pro Gly Ala Asp Met Ser Ile Tyr
510 515 520
ttc cct tac acg gaa gag aag cgg agg ttg aag aag ttc cac ccg gag 1697
Phe Pro Tyr Thr Glu Glu Lys Arg Arg Leu Lys Lys Phe His Pro Glu
525 530 535
att gaa gag ctt ctt tac agc cct gtt gag aat aca gag cac tta tgt 1745
Ile Glu Glu Leu Leu Tyr Ser Pro Val Glu Asn Thr Glu His Leu Cys
540 545 550
gta cta aaa gac cgc aac aag ccg att ctg ttt acc atg gca agg ttg 1793
Val Leu Lys Asp Arg Asn Lys Pro Ile Leu Phe Thr Met Ala Arg Leu
555 560 565 570
gac cga gtg aag aac tta act ggg ctc gta gag ttc tat gcc aag aac 1841
Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn
575 580 585
agc cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt gga gat agg 1889
Ser Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly Gly Asp Arg
590 595 600
aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg aag aag atg 1937
Arg Lys Glu Ser Lys Asp Leu Glu Glu Lys Ala Glu Met Lys Lys Met
605 610 615
tat gaa ctt atc gaa aaa tac aag ttg aat gga caa ttc aga tgg ata 1985
Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Phe Arg Trp Ile
620 625 630
tcg tcc cag atg aac cga gtg aga aat ggt gaa ctc tat cgt tat att 2033
Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr Arg Tyr Ile
635 640 645 650
tgc gac acc aag gga gca ttc gtt cag cct cct ata tac gag gct ttt 2081
Cys Asp Thr Lys Gly Ala Phe Val Gln Pro Pro Ile Tyr Glu Ala Phe
655 660 665
ggc ttg act gtt gtt gag gca atg acc tgt gga cta cca aca ttt gca 2129
Gly Leu Thr Val Val Glu Ala Met Thr Cys Gly Leu Pro Thr Phe Ala
670 675 680
aca tgt tac ggg ggc cct gct gag att ata gtt cac gga aaa tcg ggg 2177
Thr Cys Tyr Gly Gly Pro Ala Glu Ile Ile Val His Gly Lys Ser Gly
685 690 695
ttc aac atc gat cct tat aac ggt gat ttg gct gcc gag acc ctt gcc 2225
Phe Asn Ile Asp Pro Tyr Asn Gly Asp Leu Ala Ala Glu Thr Leu Ala
700 705 710
aat ttc ttc gag aag tgc aaa gcg gat cca tct tat tgg gat gag atc 2273
Asn Phe Phe Glu Lys Cys Lys Ala Asp Pro Ser Tyr Trp Asp Glu Ile
715 720 725 730
tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca tgg cag att 2321
Ser Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr Trp Gln Ile
735 740 745
tac tcc gag aag cta ttg act ctc acc gga gtt tat ggc ttt tcg aaa 2369
Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys
750 755 760
cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac att gaa atg 2417
His Val Ala Tyr Gln Glu Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met
765 770 775
ttg cat gca tgg atg tat aac aat cgg gtc aag act gtt cca cta gct 2465
Leu His Ala Trp Met Tyr Asn Asn Arg Val Lys Thr Val Pro Leu Ala
780 785 790
gtt gag taa agaaaaagca aaaggctgct tggaaaccaa ggaatttcca 2514
Val Glu
795
agaatgattg ctattgaagt tttatttttg cttctttttt tcaatttcgt aaatgtttaa 2574
tttcaacttt ggggtttccc cttggaatgt tttgaactct gggtttggtc ccattgattg 2634
c 2635
<210>2
<211>796
<212>PRT
＜213〉upland cotton
<400>2
Met Ala Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Ala Thr
1 5 10 15
His Pro Gln Gln Ala Lys Ser Ile Leu Ser Arg Ile Glu Ser Leu Gly
20 25 30
Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp Lys Glu
35 40 45
Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Gly Val Leu Arg Ser
50 55 60
Thr Gln Glu Ala Val Val Ser Ser Pro Leu Val Ala Leu Ala Ile Arg
65 70 75 80
Ser Ala Pro Gly Val Trp Glu Tyr Ile Ala Val Glu Val Gln Lys Leu
85 90 95
Phe Val Glu Glu Met Pro Val Ala Glu Tyr Leu Arg Leu Lys Glu Glu
100 105 110
Leu Val Asp Gly Ser Ser Asn Gly Glu Phe Met Leu Glu Leu Asp Phe
115 120 125
Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser Ile
130 135 140
Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu Phe
145 150 155 160
Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Glu Phe Leu Gln Ile His
165 170 175
Cys Gln Lys Gly Lys Gly Met Leu Leu Asn Asp Arg Ile Gln Asp Val
180 185 190
Asn Ser Leu Gln His Ala Leu Arg Lys Ala Glu Glu Tyr Leu Thr Pro
195 200 205
Leu Ser Ser Asp Thr Pro Tyr Ser Val Phe Glu Lys Arg Phe Leu Gly
210 215 220
Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Ala Glu His Val Leu Glu
225 230 235 240
Met Ile His Leu Leu Leu Asp Leu Leu Gln Ala Pro Asp Pro Val Ala
245 250 255
Leu Glu Ser Phe Leu Gly Arg Ile Pro Leu Val Ala Asn Val Val Ile
260 265 270
Met Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu Gly Tyr Pro
275 280 285
Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Ala Leu
290 295 300
Glu Glu Glu Leu Leu His Arg Phe Lys Leu Gln Gly Leu Asp Ile Thr
305 310 315 320
Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Ala Val Gly Thr
325 330 335
Thr Cys Gly Gln Arg Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp
340 345 350
Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp
355 360 365
Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp
370 375 380
Val Ala Ala Glu Ile Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile
385 390 395 400
Val Gly Asn Tyr Ser Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His
405 410 415
Lys Phe Asp Val Thr Gln Cys Thr Ile Ala Arg Ala Leu Glu Lys Thr
420 425 430
Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys Gln Leu Glu Asp Lys Tyr
435 440 445
His Phe Ser Cys Gln Phe Thr Ala Asp Leu Ile Ala Met Asn His Thr
450 455 460
Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu Ile Ala Gly Ser Lys Asp
465 470 475 480
Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro Gly Leu
485 490 495
Tyr Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe Asn Ile
500 505 510
Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr Glu Glu
515 520 525
Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu Leu Tyr
530 535 540
Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn
545 550 555 560
Lys Pro Ile Leu Phe Thr Met Ala Arg Leu Asp Arg Val Lys Asn Leu
565 570 575
Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu
580 585 590
Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp
595 600 605
Leu Glu Glu Lys Ala Glu Met Lys Lys Met Tyr Glu Leu Ile Glu Lys
610 615 620
Tyr Lys Leu Asn Gly Gln Phe Arg Trp Ile Ser Ser Gln Met Asn Arg
625 630 635 640
Val Arg Asn Gly Glu Leu Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Ala
645 650 655
Phe Val Gln Pro Pro Ile Tyr Glu Ala Phe Gly Leu Thr Val Val Glu
660 665 670
A1a Met Thr Cys Gly Leu Pro Thr Phe Ala Thr Cys Tyr Gly Gly Pro
675 680 685
Ala Glu Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Tyr
690 695 700
Asn Gly Asp Leu Ala Ala Glu Thr Leu Ala Asn Phe Phe Glu Lys Cys
705 710 715 720
Lys Ala Asp Pro Ser Tyr Trp Asp Glu Ile Ser Gln Gly Gly Leu Lys
725 730 735
Arg Ile Gln Glu Lys Tyr Thr Trp Gln Ile Tyr Ser Glu Lys Leu Leu
740 745 750
Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Ala Tyr Gln Glu
755 760 765
Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Leu His Ala Trp Met Tyr
770 775 780
Asn Asn Arg Val Lys Thr Val Pro Leu Ala Val Glu
785 790 795
<210>3
<211>796
<212>PRT
＜213〉upland cotton
<400>3
Met Ala Ser I1e Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Ala Thr
1 5 10 15
His Pro Gln Gln Ala Lys Ser Ile Leu Ser Arg Ile Glu Ser Leu Gly
20 25 30
Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp Lys Glu
35 40 45
Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu Arg Ser
50 55 60
Thr Gln Glu A1a Val Val Ser Ser Pro Leu Val Ala Leu Ala Ile Arg
65 70 75 80
Ser Ala Pro Gly Val Trp Glu Tyr Ile Ala Val Glu Val G1n Lys Leu
85 90 95
Phe Val G1u G1u Met Pro Val Ala Glu Tyr Leu Arg Leu Lys Glu Glu
100 105 110
Leu Val Asp Gly Ser Ser Asn Gly Glu Phe Met Leu Glu Leu Asp Phe
115 120 125
Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser Ile
130 135 140
Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu Phe
145 150 155 160
Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Glu Phe Leu Gln Ile His
165 170 175
Cys Gln Lys Gly Lys Gly Met Leu Leu Asn Asp Arg Ile Gln Asp Val
180 185 190
Asn Ser Leu Gln His Ala Leu Arg Lys Ala Glu Glu Tyr Leu Thr Pro
195 200 205
Leu Ser Ser Asp Thr Pro Tyr Ser Val Phe Glu Lys Arg Phe Leu Gly
210 215 220
Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Ala Glu His Val Leu Glu
225 230 235 240
Met Ile His Leu Leu Leu Asp Leu Leu Gln Ala Pro Asp Pro Val Ala
245 250 255
Leu Glu Ser Phe Leu Gly Arg Ile Pro Leu Val A1a Asn Val Val Ile
260 265 270
Met Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu Gly Tyr Pro
275 280 285
Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Ala Leu
290 295 300
Glu Glu Glu Leu Leu His Arg Phe Lys Leu Gln Gly Leu Asp Ile Thr
305 310 315 320
Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Ala Val Gly Thr
325 330 335
Thr Cys Gly Gln Arg Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp
340 345 350
Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp
355 360 365
Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp
370 375 380
Val Ala Ala Glu Ile Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile
385 390 395 400
Val Gly Asn Tyr Ser Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His
405 410 415
Lys Phe Asp Val Thr Gln Cys Thr Ile Ala His Ala Leu Glu Lys Thr
420 425 430
Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys Gln Leu Glu Asp Lys Tyr
435 440 445
His Phe Ser Cys Gln Phe Thr Ala Asp Leu Ile Ala Met Asn His Thr
450 455 460
Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu Ile Ala Gly Ser Lys Asp
465 470 475 480
Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro Gly Leu
485 490 495
Tyr Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe Asn Ile
500 505 510
Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr Glu Glu
515 520 525
Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu Leu Tyr
530 535 540
Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn
545 550 555 560
Lys Pro Ile Leu Phe Thr Met Ala Arg Leu Asp Arg Val Lys Asn Leu
565 570 575
Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu
580 585 590
Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp
595 600 605
Leu Glu Glu Lys Ala Glu Met Lys Lys Met Tyr Glu Leu Ile Glu Lys
610 615 620
Tyr Lys Leu Asn Gly Gln Phe Arg Trp Ile Ser Ser Gln Met Asn Arg
625 630 635 640
Val Arg Asn Gly Glu Leu Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Ala
645 650 655
Phe Val Gln Pro Pro Ile Tyr Glu Ala Phe Gly Leu Thr Val Val Glu
660 665 670
Ala Met Thr Cys Gly Leu Pro Thr Phe Ala Thr Cys Tyr Gly Gly Pro
675 680 685
Ala Glu Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Tyr
690 695 700
Asn Gly Asp Leu Ala Ala Glu Thr Leu Ala Asn Phe Phe Glu Lys Cys
705 710 715 720
Lys Ala Asp Pro Ser Tyr Trp Asp Glu Ile Ser Gln Gly Gly Leu Lys
725 730 735
Arg Ile Gln Glu Lys Tyr Thr Trp Gln Ile Tyr Ser Glu Lys Leu Leu
740 745 750
Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Ala Tyr Gln Glu
755 760 765
Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Leu His Ala Trp Met Tyr
770 775 780
Asn Asn Arg Val Lys Thr Val Pro Leu Ala Val Glu
785 790 795
<210>4
<211>3353
<212>DNA
＜213〉upland cotton
<220>
<221>misc_feature
<222>(32)..(34)
＜223〉initiator codon
<220>
＜221〉exon
<222>(32)..(111)
＜223〉exons 1
<220>
＜221〉exon
<222>(197)..(314)
＜223〉exon 2
<220>
＜221〉exon
<222>(394)..(545)
＜223〉exon 3
<220>
＜221〉exon
<222>(625)..(817)
＜223〉exon 4
<220>
＜221〉exon
<222>(897)..(1328)
＜223〉exon 5
<220>
＜221〉exon
<222>(1399)..(1689)
＜223〉exon 6
<220>
＜221〉exon
<222>(1783)..(1949)
＜223〉exon 7
<220>
＜221〉exon
<222>(2050)..(2274)
＜223〉exon 8
<220>
＜221〉exon
<222>(2358)..(3060)
＜223〉exon 9
<220>
＜221〉exon
<222>(3169)..(3195)
＜223〉exons 10
<220>
<221>misc_feature
<222>(3196)..(3171)
＜223〉terminator codon
<400>4
cccttctgcc atttcaggaa cccaaaaaac a atg gct tca atc agt gtt tgt 52
Met Ala Ser Ile Ser Val Cys
1 5
gag cgt ttg ggt gaa tct cta gct act cat cca cag cag gca aag tct 100
Glu Arg Leu Gly Glu Ser Leu Ala Thr His Pro Gln Gln Ala Lys Ser
10 15 20
atc ttg tca ag gtataatgta aataagtgct gccttcaaat ttctggtctt 151
Ile Leu Ser Arg
25
tcttcgttat tttttattac attgtttata tatgtcatat tgcag g att gaa agc 206
Ile Glu Ser
30
ctc gga aag ggt att cat aag tct caa aag ctg ctc tcg gtt ctc gat 254
Leu Gly Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp
35 40 45
aaa gag gcc gga aat caa gca ctt gat ggg atg gtg gtg gag gtc ctc 302
Lys Glu Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu
50 55 60
agg tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtttcatg 354
Arg Ser Thr Gln
65
atgagaatat tggactgata tgttaccgtt atctaatag gaa gct gta gtg tcg 408
Glu Ala Val Val Ser
70
tct cca ttg gtt gcc ctt gcc att cgt tca gct cct gga gtt tgg gag 456
Ser Pro Leu Val Ala Leu Ala Ile Arg Ser Ala Pro Gly Val Trp Glu
75 80 85
tac att gct gtg gag gtc caa aag ctt ttt gtg gag gaa atg ccc gtt 504
Tyr Ile Ala Val Glu Val Gln Lys Leu Phe Val Glu Glu Met Pro Val
90 95 100
gct gag tat cta cgg ttg aag gaa gaa ctt gtt gat gga ag 545
Ala Glu Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser
105 110 115
gtaggtagtt ttctcttaca gctcctggtt tatattatgt ttgagtttat gttgaacaat 605
gctcaatgat gtggttcag c tcc aat ggc gag ttt atg ttg gaa ttg gac 655
Ser Asn Gly Glu Phe Met Leu Glu Leu Asp
120 125
ttt ggt gca ttc aat aat tct gtt cct cgt cca tct ctt tca aag tcc 703
Phe Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser
130 135 140
att ggt aat ggc atg gac ttc ctc aac cgc cac ctt tct gcc aag cta 751
Ile Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu
145 150 155
ttt caa gac aag gag aac ttg aac ttg ttg ctt gaa ttt ctc caa att 799
Phe Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Glu Phe Leu Gln Ile
160 165 170 175
cac tgc cag aag gga aag gtaaaagttt cactttcact atttgttttt 847
His Cys Gln Lys Gly Lys
180
cctttctgta tacatcaaaa cttaagaatc gttcaatgtt atgccacag ggt atg ctg 905
Gly Met Leu
ttg aat gac aga atc caa gat gtg aat tcc ctc caa cat gca tta agg 953
Leu Asn Asp Arg Ile Gln Asp Val Asn Ser Leu Gln His Ala Leu Arg
185 190 195 200
aag gcc gag gag tat ctg act cct cta tcc tcg gat acc ccg tac tca 1001
Lys Ala Glu Glu Tyr Leu Thr Pro Leu Ser Ser Asp Thr Pro Tyr Ser
205 210 215
gtt ttc gag aaa agg ttt ctg ggg att ggt ttg gag aag gga tgg ggt 1049
Val Phe Glu Lys Arg Phe Leu Gly Ile Gly Leu Glu Lys Gly Trp Gly
220 225 230
gat aat gct gag cat gtt ctt gag atg atc cat ctt cta ttg gat ctc 1097
Asp Asn Ala Glu His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu
235 240 245
ctt cag gca cct gat cct gtc gca ctt gaa agc ttc ctt gga aga atc 1145
Leu Gln Ala Pro Asp Pro Val Ala Leu Glu Ser Phe Leu Gly Arg Ile
250 255 260
cca ttg gtc gcc aat gtt gta atc atg act ccc cat gga tac ttt gcc 1193
Pro Leu Val Ala Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Ala
265 270 275 280
caa gac aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat 1241
Gln Asp Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr
285 290 295
atc tta gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt ttc 1289
Ile Leu Asp Gln Val Arg Ala Leu Glu Glu Glu Leu Leu His Arg Phe
300 305 310
aag ctg caa gga ctc gac att acc cca cgt atc cta gtc gtgagtatct 1338
Lys Leu Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val
315 320 325
gaatgttaat tcagtttcga tcattatgtt taatgcttaa tttggctgtt tgtgtaccag 1398
att act cgg ctc ctc cct gat gct gta gga aca act tgc ggt cag cgt 1446
Ile Thr Arg Leu Leu Pro Asp Ala Val Gly Thr Thr Cys Gly Gln Arg
330 335 340
ctt gag aaa gtg tat gga acc aag tat tct gat att ctt cgg gta ccc 1494
Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp Ile Leu Arg Val Pro
345 350 355
ttc aga act gag aag gga att gta cgt cca tgg atc tca cga ttc aaa 1542
Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp Ile Ser Arg Phe Lys
360 365 370
gtc tgg cct tac ctg gag act tac act aag gat gtt gca gct gag atc 1590
Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp Val Ala Ala Glu Ile
375 380 385
acc aaa gag ttc cag ggc aag cct gat ctg att gtt ggg aac tac agt 1638
Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile Val Gly Asn Tyr Ser
390 395 400 405
gat gga aac att gtt gct tct tta ttg gca cat aag ttt gat gtt aca 1686
Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His Lys Phe Asp Val Thr
410 415 420
cag gtttgttaaa taccaattaa ttgaacaagt tccttgctgt attttgatgt 1739
Gln
tccccgtgca atctaaaaac ttggttttca atgttttatg cag tgc act att gct 1794
Cys Thr Ile Ala
425
cat gca ctc gag aag acg aaa tac ccg gat tca gac att aac tgg aaa 1842
His Ala Leu Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys
430 435 440
cag ctt gag gat aag tat cac ttc tcc tgt cag ttt act gct gat ctt 1890
Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Ala Asp Leu
445 450 455
att gct atg aac cat act gat ttt atc atc acc agc acc ttc caa gag 1938
Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu
460 465 470
att gct gga ag gtaatctatc tcccacttta ctacatttgg ttcccttgta 1989
Ile Ala Gly Ser
475
cgttgtaaat gttgccacct atttgagtcc ctaacaatgt tttcattcca tgattttcag 2049
c aag gac act ctc ggc caa tac gag agt cac att gct ttc act ctt cca 2098
Lys Asp Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro
480 485 490
ggg ctc tac cgc gtt gtt gat ggg atc gat gtt ttt gat ccc aaa ttc 2146
Gly Leu Tyr Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe
495 500 505 510
aat att gtc tcc cct ggt gct gat atg agc ata tac ttc cct tac acg 2194
Asn Ile Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr
515 520 525
gaa gag aag cgg agg ttg aag aag ttc cac ccg gag att gaa gag ctt 2242
Glu Glu Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu
530 535 540
ctt tac agc cct gtt gag aat aca gag cac tt gtgagatttt ggtcctattt 2294
Leu Tyr Ser Pro Val Glu Asn Thr Glu His Leu
545 550
ttacaccctt ttcactagat tttgtgttca gtgttaacaa tgtcttaaat tgttgtctca 2354
cag a tgt gta cta aaa gac cgc aac aag ccg att ctg ttt acc atg gca 2403
Cys Val Leu Lys Asp Arg Asn Lys Pro Ile Leu Phe Thr Met Ala
555 560 565
agg ttg gac cga gtg aag aac tta act ggg ctc gta gag ttc tat gcc 2451
Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Phe Tyr Ala
570 575 580
aag aac agc cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt gga 2499
Lys Asn Ser Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly Gly
585 590 595 600
gat agg aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg aag 2547
Asp Arg Arg Lys Glu Ser Lys Asp Leu Glu Glu Lys Ala Glu Met Lys
605 610 615
aag atg tat gaa ctt atc gaa aaa tac aag ttg aat gga caa ttc aga 2595
Lys Met Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Phe Arg
620 625 630
tgg ata tcg tcc cag atg aac cga gtg aga aat ggt gaa ctc tat cgt 2643
Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr Arg
635 640 645
tat att tgc gac acc aag gga gca ttc gtt cag cct cct ata tac gag 2691
Tyr Ile Cys Asp Thr Lys Gly Ala Phe Val Gln Pro Pro Ile Tyr Glu
650 655 660
gct ttt ggc ttg act gtt gtt gag gca atg acc tgt gga cta cca aca 2739
Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr Cys Gly Leu Pro Thr
665 670 675 680
ttt gca aca tgt tac ggg ggc cct gct gag att ata gtt cac gga aaa 2787
Phe Ala Thr Cys Tyr Gly Gly Pro Ala Glu Ile Ile Val His Gly Lys
685 690 695
tcg ggg ttc aac atc gat cct tat aac ggt gat ttg gct gcc gag acc 2835
Ser Gly Phe Asn Ile Asp Pro Tyr Asn Gly Asp Leu Ala Ala Glu Thr
700 705 710
ctt gcc aat ttc ttc gag aag tgc aaa gcg gat cca tct tat tgg gat 2883
Leu Ala Asn Phe Phe Glu Lys Cys Lys Ala Asp Pro Ser Tyr Trp Asp
715 720 725
gag atc tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca tgg 2931
Glu Ile Ser Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr Trp
730 735 740
cag att tac tcc gag aag cta ttg act ctc acc gga gtt tat ggc ttt 2979
Gln Ile Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly Phe
745 750 755 760
tcg aaa cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac att 3027
Ser Lys His Val Ala Tyr Gln Glu Gln Arg Gly Arg Lys Arg Tyr Ile
765 770 775
gaa atg ttg cat gca tgg atg tat aac aat cgg gtaaaacaat catgtctatt 3080
Glu Met Leu His Ala Trp Met Tyr Asn Asn Arg
780 785
attattatta catcaaacat cattttactc tttcattgaa ctgaaatttt catgtttctg 3140
aatgtttgtt taattgtgtg ctttacag gtc aag act gtt cca cta gct gtt 3192
Val Lys Thr Val Pro Leu Ala Val
790 795
gag taaagaaaaa gcaaaaggct gcttggaaac caaggaattt ccaagaatga 3245
Glu
ttgctattga agttttattt ttgcttcttt ttttcaattt cgtaaatgtt taatttcaac 3305
tttggggttt ccccttggaa tgttttgaac tctgggtttg gtcccatt 3353
<210>5
<211>796
<212>PRT
＜213〉sea island cotton (Gossypium barbadense)
<400>5
Met Ala Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Ala Thr
1 5 10 15
His Pro Gln Gln Ala Lys Ser Ile Leu Ser Arg Ile Glu Ser Leu Gly
20 25 30
Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp Lys Glu
35 40 45
Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu Arg Ser
50 55 60
Thr Gln Glu Ala Val Val Ser Ser Pro Leu Val Ala Leu Ala Ile Arg
65 70 75 80
Ser Ala Pro Gly Val Trp Glu Tyr Ile Ala Val Glu Val Gln Lys Leu
85 90 95
Phe Val Glu Glu Met Pro Val Ala Glu Tyr Leu Arg Leu Lys Glu Glu
100 105 110
Leu Val Asp Gly Ser Ser Asn Gly Glu Phe Met Leu Glu Leu Asp Phe
115 120 125
Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser Ile
130 135 140
Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu Phe
145 150 155 160
Gln Asp Lys Glu Ser Leu Asn Leu Leu Leu Glu Phe Leu Gln Ile His
165 170 175
Cys Gln Lys Gly Lys Gly Met Leu Leu Asn Asp Arg Ile Gln Asp Val
180 185 190
Asn Ser Leu Gln His Ala Leu Arg Lys Ala Glu Glu Tyr Leu Thr Pro
195 200 205
Leu Ser Ser Asp Thr Pro Tyr Ser Val Phe Glu Lys Arg Phe Leu Gly
210 215 220
Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Ala Glu His Val Leu Glu
225 230 235 240
Met Ile His Leu Leu Leu Asp Leu Leu Gln Ala Pro Asp Pro Val Ala
245 250 255
Leu Glu Ser Phe Leu Gly Arg Ile Pro Leu Val Ala Asn Val Val Ile
260 265 270
Met Thr Pro His Gly Tyr Phe Ala Gln Tyr Asn Val Leu Gly Tyr Pro
275 280 285
Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Ala Leu
290 295 300
Glu Glu Glu Leu Leu His Arg Phe Lys Leu Gln Gly Leu Asp Ile Thr
305 310 315 320
Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Ala Val Gly Thr
325 330 335
Thr Cys Gly Gln Arg Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp
340 345 350
Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp
355 360 365
Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp
370 375 380
Val Ala Ala Glu Ile Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile
385 390 395 400
Val Gly Asn Tyr Ser Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His
405 410 415
Lys Phe Asp Val Thr Gln Cys Thr Ile Ala His Ala Leu Glu Lys Thr
420 425 430
Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys Gln Leu Glu Asp Lys Tyr
435 440 445
His Phe Ser Cys Gln Phe Thr Ala Asp Leu Ile Ala Met Asn His Thr
450 455 460
Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu Ile Ala Gly Ser Lys Asp
465 470 475 480
Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro Gly Leu
485 490 495
Tyr Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe Asn Ile
500 505 510
Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr Glu Glu
515 520 525
Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu Leu Tyr
530 535 540
Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn
545 550 555 560
Lys Pro Ile Leu Phe Thr Met Ala Arg Leu Asp Arg Val Lys Asn Leu
565 570 575
Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu
580 585 590
Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp
595 600 605
Leu Glu Glu Lys Ala Glu Met Lys Lys Met Tyr Glu Leu Ile Glu Lys
610 615 620
Tyr Lys Leu Asn Gly Gln Phe Arg Trp Ile Ser Ser Gln Met Asn Arg
625 630 635 640
Val Arg Asn Gly Glu Leu Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Ala
645 650 655
Phe Val Gln Pro Ala Ile Tyr Glu Ala Phe Gly Leu Thr Val Val Glu
660 665 670
Ala Met Thr Cys Gly Leu Pro Thr Phe Ala Thr Cys Tyr Gly Gly Pro
675 680 685
Ala Glu Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Tyr
690 695 700
Asn Gly Asp Leu Ala Ala Glu Thr Leu Ala Asn Phe Phe Glu Lys Cys
705 710 715 720
Lys Ala Asp Pro Ser Tyr Trp Asp Glu Ile Ser Gln Gly Gly Leu Lys
725 730 735
Arg Ile Gln Glu Lys Tyr Thr Trp Gln Ile Tyr Ser Glu Lys Leu Leu
740 745 750
Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Ala Tyr Gln Glu
755 760 765
Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Leu His Ala Trp Met Tyr
770 775 780
Asn Asn Arg Val Lys Thr Val Pro Leu Ala Val Glu
785 790 795
<210>6
<211>3352
<212>DNA
＜213〉sea island cotton
<220>
<221>misc_feature
<222>(32)..(34)
＜223〉initiator codon
<220>
＜221〉exon
<222>(32)..(111)
＜223〉exons 1
<220>
＜221〉exon
<222>(197)..(314)
＜223〉exon 2
<220>
＜221〉exon
<222>(394)..(545)
＜223〉exon 3
<220>
＜221〉exon
<222>(625)..(817)
＜223〉exon 4
<220>
＜221〉exon
<222>(897)..(1328)
＜223〉exon 5
<220>
＜221〉exon
<222>(1399)..(1689)
＜223〉exon 6
<220>
＜221〉exon
<222>(1783)..(1949)
＜223〉exon 7
<220>
＜221〉exon
<222>(2050)..(2274)
＜223〉exon 8
<220>
＜221〉exon
<222>(2357)..(3059)
＜223〉exon 9
<220>
＜221〉exon
<222>(3168)..(3194)
＜223〉exons 10
<220>
<221>misc_feature
<222>(3195)..(3197)
＜223〉terminator codon
<400>6
cccttctgcc atttcaggaa cccaaaaaac a atg gct tca atc agt gtt tgt 52
Met Ala Ser Ile Ser Val Cys
1 5
gag cgt ttg ggt gaa tct tta gct act cat cca cag cag gca aag tct 100
Glu Arg Leu Gly Glu Ser Leu Ala Thr His Pro Gln Gln Ala Lys Ser
10 15 20
atcttg tca ag gtataatgta aataagtgct gccttcaaat ttctggtctt 151
Ile Leu Ser Arg
25
tcttcgttat tttttattac attgtttata tatgtcatat tgcag g att gaa agc 206
Ile Glu Ser
30
ctc gga aag ggt att cat aag tct caa aag ctg ctc tcg gtt ctc gat 254
Leu Gly Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp
35 40 45
aaa gag gcc gga aat caa gca ctt gat ggg atg gtg gtg gag gtc ctc 302
Lys Glu Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu
50 55 60
agg tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtttcatg 354
Arg Ser Thr Gln
65
atgagaatat tggactgata tgttaccgtt atctaatag gaa gct gta gtg tcg 408
Glu Ala Val Val Ser
70
tct cca ttg gtt gcc ctt gcc att cgt tca gct cct gga gtt tgg gag 456
Ser Pro Leu Val Ala Leu Ala Ile Arg Ser Ala Pro Gly Val Trp Glu
75 80 85
tac att gct gtg gag gtc caa aag ctt ttt gtg gag gaa atg ccc gtt 504
Tyr Ile Ala Val Glu Val Gln Lys Leu Phe Val Glu Glu Met Pro Val
90 95 100
gct gag tat cta cgg ttg aag gaa gaa ctt gtt gat gga ag 545
Ala Glu Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser
105 110 115
gtaggtagtt ttctcttaca gctcctggtt tatgttatgt ttgagtttat gttgaacaat 605
gctcaatgat gtggttcag c tcc aat ggc gag ttt atg ttg gaa ttg gac 655
Ser Asn Gly Glu Phe Met Leu Glu Leu Asp
120 125
ttt ggt gca ttc aat aat tct gtt cct cgt cca tct ctt tca aag tcc 703
Phe Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser
130 135 140
att ggt aat ggc atg gac ttc ctc aac cgc cac ctt tct gcc aag cta 751
Ile Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu
145 150 155
ttt caa gac aag gag agc ttg aac ttg ttg ctt gaa ttt ctc caa att 799
Phe Gln Asp Lys Glu Ser Leu Asn Leu Leu Leu Glu Phe Leu Gln Ile
160 165 170 175
cac tgc cag aag gga aag gtaaaagttt cactttcact atttgttttt 847
His Cys Gln Lys Gly Lys
180
cctttctgta tacatcaaaa cttaagaatc gttcaatgtt atgccacag ggt atg ctg 905
Gly Met Leu
ttg aat gac aga atc caa gat gtg aat tcc ctc caa cat gca tta agg 953
Leu Asn Asp Arg Ile Gln Asp Val Asn Ser Leu Gln His Ala Leu Arg
185 190 195 200
aag gcc gag gag tat ctg act cct cta tcc tcg gat acc ccg tac tca 1001
Lys Ala Glu Glu Tyr Leu Thr Pro Leu Ser Ser Asp Thr Pro Tyr Ser
205 210 215
gtt ttc gag aaa agg ttt ctg ggg att ggt ttg gag aag gga tgg ggt 1049
Val Phe Glu Lys Arg Phe Leu Gly Ile Gly Leu Glu Lys Gly Trp Gly
220 225 230
gat aat gct gag cat gtt ctt gag atg atc cat ctt cta ttg gat ctc 1097
Asp Asn Ala Glu His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu
235 240 245
ctt cag gca cct gat cct gtc gca ctt gaa agc ttc ctt gga aga atc 1145
Leu Gln Ala Pro Asp Pro Val Ala Leu Glu Ser Phe Leu Gly Arg Ile
250 255 260
cca ttg gtc gcc aat gtt gta atc atg act ccc cat gga tac ttt gcc 1193
Pro Leu Val Ala Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Ala
265 270 275 280
caa tac aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat 1241
Gln Tyr Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr
285 290 295
atc tta gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt ttc 1289
Ile Leu Asp Gln Val Arg Ala Leu Glu Glu Glu Leu Leu His Arg Phe
300 305 310
aag ctg caa gga ctc gac att acc cca cgt atc cta gtc gtgagtatct 1338
Lys Leu Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val
315 320 325
gaatgttaat tcagtttcga tcattatgtt taatgcttaa tttggctgtt tgtgtaccag 1398
att act cgg ctc ctc cct gat gct gta gga aca act tgc ggt cag cgt 1446
lle Thr Arg Leu Leu Pro Asp Ala Val Gly Thr Thr Cys Gly Gln Arg
330 335 340
ctt gag aaa gtg tat gga acc aag tat tct gat att ctt cgg gta ccc 1494
Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp Ile Leu Arg Val Pro
345 350 355
ttc aga act gag aag gga att gta cgt cca tgg atc tca cga ttc aaa 1542
Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp Ile Ser Arg Phe Lys
360 365 370
gtc tgg cct tac ctg gag act tac act aag gat gtt gca gct gag atc 1590
Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp Val Ala Ala Glu Tle
375 380 385
acc aaa gag ttc cag ggc aag cct gat ctg att gtt ggg aac tac agt 1638
Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile Val Gly Asn Tyr Ser
390 395 400 405
gat gga aac att gtt gct tct tta ttg gca cat aag ttt gat gtt aca 1686
Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His Lys Phe Asp Val Thr
410 415 420
caggtttgttaaa taccaattaa ttgaacaagt tccttgctgt attttgatgt 1739
Gln
tccccgtgca atctaaaaac ttggttttca atgttttatg cag tgc act att gct 1794
Cys Thr Ile Ala
425
cat gca ctc gag aag acg aaa tac ccg gat tca gac att aac tgg aaa 1842
His Ala Leu Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys
430 435 440
cag ctt gag gat aag tat cac ttc tcc tgt cag ttt act gct gat ctt 1890
Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Ala Asp Leu
445 450 455
att gct atg aac cat act gat ttt atc atc acc agc acc ttc caa gag 1938
Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu
460 465 470
att gct gga ag gtaatctatc tcccacttta ctacatttgg ttcccttgta 1989
Ile Ala Gly Ser
475
cgttgtaaat gttgccacct atttgagtcc ctaacaatgt tttcattcca tgattttcag 2049
c aag gac act ctc ggc caa tac gag agt cac att gct ttc act ctt cca 2098
Lys Asp Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro
480 485 490
ggg ctc tac cgc gtt gtt gat ggg atc gat gtt ttt gat ccc aaa ttc 2146
Gly Leu Tyr Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe
495 500 505 510
aat att gtc tcc cct ggt gct gat atg agc ata tac ttc cct tac acg 2194
Asn Ile Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr
515 520 525
gaa gag aag cgg agg ttg aag aag ttc cac ccg gag att gaa gag ctt 2242
Glu Glu Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu
530 535 540
ctt tac agc cct gtt gag aat aca gag cac tt gtgagatttt ggtcctattt 2294
Leu Tyr Ser Pro Val Glu Asn Thr Glu His Leu
545 550
ttacaccctt tcactagatt ttgtgttcag tgttaacaat gtcttaaatt gttgtctcac 2354
ag a tgt gta cta aaa gac cgc aac aag ccg att ctg ttt acc atg gca 2402
Cys Val Leu Lys Asp Arg Asn Lys Pro Ile Leu Phe Thr Met Ala
555 560 565
agg ttg gac cga gtg aag aac tta act ggg ctc gta gag ttc tat gcc 2450
Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Phe Tyr Ala
570 575 580
aag aac agc cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt gga 2498
Lys Asn Ser Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly Gly
585 590 595 600
gat agg aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg aag 2546
Asp Arg Arg Lys Glu Ser Lys Asp Leu Glu Glu Lys Ala Glu Met Lys
605 610 615
aag atg tat gaa ctt atc gaa aaa tac aag ttg aat gga caa ttc aga 2594
Lys Met Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Phe Arg
620 625 630
tgg ata tcg tcc cag atg aac cga gtg aga aat ggt gaa ctc tat cgt 2642
Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr Arg
635 640 645
tat att tgc gac acc aag gga gca ttc gtt cag cct gct ata tac gag 2690
Tyr Ile Cys Asp Thr Lys Gly Ala Phe Val Gln Pro Ala Ile Tyr Glu
650 655 660
gct ttt ggc ttg act gtt gtt gag gca atg acc tgt gga cta cca aca 2738
Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr Cys Gly Leu Pro Thr
665 670 675 680
ttt gca aca tgt tac ggg ggc cct gct gag att ata gtt cac gga aaa 2786
Phe Ala Thr Cys Tyr Gly Gly Pro Ala Glu Ile Ile Val His Gly Lys
685 690 695
tcg ggg ttc aac atc gat cct tat aac ggt gat ttg gct gcc gag acc 2834
Ser Gly Phe Asn Ile Asp Pro Tyr Asn Gly Asp Leu Ala Ala Glu Thr
700 705 710
ctt gcc aat ttc ttc gag aag tgc aaa gcg gat cca tct tat tgg gat 2882
Leu Ala Asn Phe Phe Glu Lys Cys Lys Ala Asp Pro Ser Tyr Trp Asp
715 720 725
gag atc tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca tgg 2930
Glu Ile Ser Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr Trp
730 735 740
cag att tac tcc gag aag cta ttg act ctc acc gga gtt tat ggc ttt 2978
Gln Ile Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly Phe
745 750 755 760
tcg aaa cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac att 3026
Ser Lys His Val Ala Tyr Gln Glu Gln Arg Gly Arg Lys Arg Tyr Ile
765 770 775
gaa atg ttg cat gca tgg atg tat aac aat cgg gtaaaacaat catgtctatt 3079
Glu Met Leu His Ala Trp Met Tyr Asn Asn Arg
780 785
attattatta catcaaacat cattttactc tttcattgaa ctgaaaattt catgtttctg 3139
aatgtttgtt taattgtgtg ctttacag gtc aag act gtt cca cta gct gtt 3191
Val Lys Thr Val Pro Leu Ala Val
790 795
gag taaagaaaaa gcaaaaggct gcttggaaac caaggaattt ccaagaatga 3244
Glu
ttgctattga agttttattt ttgcttcttt ttttcaattt ggtaaatgtt taatttcaac 3304
tttggggttt ccccttggaa tgttttgaac tctgggtttg gtcccatt 3352
<210>7
<211>796
<212>PRT
＜213〉tree cotton (Gossypium arboretum)
<400>7
Met Ala Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Ala Thr
1 5 10 15
His Pro Gln Gln Ala Lys Ser Ile Leu Ser Arg Ile Glu Ser Leu Gly
20 25 30
Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp Lys Glu
35 40 45
Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu Arg Ser
50 55 60
Thr Gln Glu Ala Val Val Ser Ser Pro Leu Val Ala Leu Ala Ile Arg
65 70 75 80
Ser Ala Pro Gly Val Trp Glu Tyr Ile Ala Val Glu Val Gln Lys Leu
85 90 95
Phe Val Glu Glu Met Pro Val Ala Glu Tyr Leu Arg Leu Lys Glu Glu
100 105 110
Leu Val Asp Gly Ser Ser Asn Gly Glu Phe Met Leu Glu Leu Asp Phe
115 120 125
Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser Ile
130 135 140
Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu Phe
145 150 155 160
Gln Asp Lys Glu Asn Leu Ser Leu Leu Leu Glu Phe Leu Gln Ile His
165 170 175
Cys Gln Lys Gly Lys Gly Met Leu Leu Asn Asp Arg Ile Gln Asp Val
180 185 190
Asn Ser Leu Gln His Ala Leu Arg Lys Ala Glu Glu Tyr Leu Thr Pro
195 200 205
Leu Thr Ser Asp Thr Pro Tyr Ser Val Phe Glu Lys Arg Phe Arg Glu
210 215 220
Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Ala Glu His Val Leu Glu
225 230 235 240
Met Ile His Leu Leu Leu Asp Leu Leu Gln Ala Pro Asp Pro Val Ala
245 250 255
Leu Glu Ser Phe Leu Gly Thr Ile Pro Leu Val Ala Asn Val Val Ile
260 265 270
Met Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu Gly Tyr Pro
275 280 285
Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Ala Leu
290 295 300
Glu Glu Glu Leu Leu His Arg Phe Lys Leu Gln Gly Leu Asp Ile Thr
305 310 315 320
Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Ala Val Gly Thr
325 330 335
Thr Cys Gly Gln Arg Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp
340 345 350
Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp
355 360 365
Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp
370 375 380
Val Ala Ala Glu Val Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile
385 390 395 400
Val Gly Asn Tyr Ser Asp Gly Asn Ile Val Ala Ser Leu Leu Ala Leu
405 410 415
Lys Phe Asp Val Thr Gln Cys Thr Ile Ala His Ala Leu Glu Lys Thr
420 425 430
Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys Gln Leu Glu Asp Lys Tyr
435 440 445
His Phe Ser Cys Gln Phe Thr Ala Asp Leu Ile Ala Met Asn His Thr
450 455 460
Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu Ile Ala Gly Ser Lys Asp
465 470 475 480
Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro Gly Leu
485 490 495
Tyr Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe Asn Ile
500 505 510
Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr Glu Glu
515 520 525
Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu Leu Tyr
530 535 540
Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn
545 550 555 560
Lys Pro Val Leu Phe Thr Met Ala Arg Leu Asp Arg Val Lys Asn Leu
565 570 575
Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu
580 585 590
Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp
595 600 605
Leu Glu Glu Lys Ala Glu Met Lys Lys Met Tyr Glu Leu Ile Glu Lys
610 615 620
Tyr Lys Leu Asn Gly Gln Leu Arg Trp Ile Ser Ser Gln Met Asn Arg
625 630 635 640
Val Arg Asn Gly Glu Leu Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Ala
645 650 655
Phe Val Gln Pro Ala Ile Tyr Glu Ala Phe Gly Leu Thr Val Val Glu
660 665 670
Ala Met Thr Cys Gly Leu Pro Thr Phe Ala Thr Cys Tyr Gly Gly Pro
675 680 685
Ala Glu Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Tyr
690 695 700
Asn Gly Asp Leu Ala Ala Glu Thr Leu Ala Asn Phe Phe Glu Lys Cys
705 710 715 720
Lys Ala Asp Pro Ser Tyr Trp Asp Glu Ile Ser Gln Gly Gly Leu Lys
725 730 735
Arg Ile Gln Glu Lys Tyr Thr Trp Gln Ile Tyr Ser Glu Lys Leu Leu
740 745 750
Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Ala Tyr Gln Glu
755 760 765
Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Leu His Ala Leu Met Tyr
770 775 780
Asn Asn Arg Val Lys Thr Val Pro Leu Ala Val Glu
785 790 795
<210>8
<211>3352
<212>DNA
＜213〉tree cotton
<220>
＜221〉exon
<222>(32)..(111)
＜223〉exons 1
<220>
<221>misc_feature
<222>(32)..(34)
＜223〉ATG initiator codon
<220>
＜221〉exon
<222>(197)..(314)
＜223〉exon 2
<220>
＜221〉exon
<222>(390)..(541)
＜223〉exon 3
<220>
＜221〉exon
<222>(621)..(813)
＜223〉exon 4
<220>
＜221〉exon
<222>(893)..(1324)
＜223〉exon 5
<220>
＜221〉exon
<222>(1395)..(1685)
＜223〉exon 6
<220>
＜221〉exon
<222>(1779)..(1945)
＜223〉exon 7
<220>
＜221〉exon
<222>(2046)..(2270)
＜223〉exon 8
<220>
＜221〉exon
<222>(2355)..(3057)
＜223〉exon 9
<220>
＜221〉exon
<222>(3162)..(3188)
＜223〉exons 10
<220>
<221>misc_feature
<222>(3189)..(3191)
＜223〉terminator codon
<400>8
cccttctgcc atttcaggaa ccaaaaaagc a atg gct tca atc agt gtc tgt 52
Met Ala Ser Ile Ser Val Cys
1 5
gag cgt ttg ggt gaa tct cta gct act cat cca cag cag gca aag tct 100
Glu Arg Leu Gly Glu Ser Leu Ala Thr His Pro Gln Gln Ala Lys Ser
10 15 20
atc ttg tca ag gtataatgta aataagtgct gccttcaaag ttctggtctt 151
Ile Leu Ser Arg
25
tctttgttat tttttgttac attgtttata tatgtcatat tgcag g att gaa agc 206
Ile Glu Ser
30
ctt gga aag ggt att cat aag tct caa aag ctg ctc tca gtt ctc gat 254
Leu Gly Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp
35 40 45
aaa gag gcc gga aat caa gca ctt gat ggg atg gtg gta gag gtc ctc 302
Lys Glu Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu
50 55 60
agg tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtatgatg 354
Arg Ser Thr Gln
65
agaatattgg actgatatgt taccattatc tatag gaa gct gta gtg tcg tct 407
Glu Ala Val Val Ser Ser
70
cca ttg gtt gcc ctt gcc att cgt tca gct cct gga gtt tgg gag tac 455
Pro Leu Val Ala Leu Ala Ile Arg Ser Ala Pro Gly Val Trp Glu Tyr
75 80 85
att gct gtg gag gtc caa aag ctt ttt gtg gag gaa atg ccc gtt gct 503
Ile Ala Val Glu Val Gln Lys Leu Phe Val Glu Glu Met Pro Val Ala
90 95 100
gag tat cta cgg ttg aag gaa gaa ctt gtt gat gga ag gtaggtagtt 551
Glu Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser
105 110 115
ttctcttaca gctcctggtt tatgttatgt ttgagtttat gttgaacaat gctcaatgat 611
gtggttcag c tcc aat ggc gag ttt atg ttg gaa ttg gac ttt ggt gca 660
Ser Asn Gly Glu Phe Met Leu Glu Leu Asp Phe Gly Ala
120 125 130
ttc aat aat tct gtt cct cgt cca tct ctt tca aag tcc att ggt aat 708
Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser Ile Gly Asn
135 140 145
ggc atg gac ttc ctc aac cgc cac ctt tct gcc aag cta ttt caa gac 756
Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu Phe Gln Asp
150 155 160
aag gag aac ttg agc ttg ttg ctt gaa ttt ctc caa att cac tgc cag 804
Lys Glu Asn Leu Ser Leu Leu Leu Glu Phe Leu Gln Ile His Cys Gln
165 170 175
aag gga aag gtaaaagttt cactttcact atttgttttt cctttctgta 853
Lys Gly Lys
180
tacatcaaaa cttaagaatc gttcaatgtt atgccacag ggt atg ctg ttg aat 907
Gly Met Leu Leu Asn
185
gac aga atc caa gat gtg aat tcc ctc caa cat gca ttg agg aag gcc 955
Asp Arg Ile Gln Asp Val Asn Ser Leu Gln His Ala Leu Arg Lys Ala
190 195 200
gag gag tat ctg act cct cta acg tcg gat acc ccg tac tcg gtt ttc 1003
Glu Glu Tyr Leu Thr Pro Leu Thr Ser Asp Thr Pro Tyr Ser Val Phe
205 210 215
gag aaa agg ttt cgg gag att ggt ttg gag aag gga tgg ggt gat aat 1051
Glu Lys Arg Phe Arg Glu Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn
220 225 230
gct gag cat gtt ctt gag atg atc cat ctt cta ttg gat ctc ctt cag 1099
Ala Glu His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu Leu Gln
235 240 245 250
gca cct gat cct gtc gca ctt gaa agc ttc ctt gga aca atc cca ttg 1147
Ala Pro Asp Pro Val Ala Leu Glu Ser Phe Leu Gly Thr Ile Pro Leu
255 260 265
gtc gcc aat gtt gta atc atg act ccc cat gga tac ttt gcc caa gac 1195
Val Ala Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Ala Gln Asp
270 275 280
aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat atc tta 1243
Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr Ile Leu
285 290 295
gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt ttc aag ctg 1291
Asp Gln Val Arg Ala Leu Glu Glu Glu Leu Leu His Arg Phe Lys Leu
300 305 310
caa gga ctc gac att acc cca cga atc cta gtc gtgagtatct gaatattaat 1344
Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val
315 32 0325
tcggtttcga tcattatgtt taatgcttaa tttggctgtt tatgtaccag att act 1400
Ile Thr
cgg ctc ctc cct gat gct gta gga aca act tgc ggt cag cgt ctt gag 1448
Arg Leu Leu Pro Asp Ala Val Gly Thr Thr Cys Gly Gln Arg Leu Glu
330 335 340
aaa gtg tat gga acc aag tat tct gat att ctt cgg gta ccc ttc aga 1496
Lys Val Tyr Gly Thr Lys Tyr Ser Asp Ile Leu Arg Val Pro Phe Arg
345 350 355
act gag aag gga att gta cgt cca tgg atc tca cga ttc aaa gtc tgg 1544
Thr Glu Lys Gly Ile Val Arg Pro Trp Ile Ser Arg Phe Lys Val Trp
360 365 370 375
cct tac ctg gag act tac act aag gat gtt gca gct gag gtc acc aaa 1592
Pro Tyr Leu Glu Thr Tyr Thr Lys Asp Val Ala Ala Glu Val Thr Lys
380 385 390
gag ttc cag ggc aaa cct gat ctg att gtt ggg aac tac agt gat gga 1640
Glu Phe Gln Gly Lys Pro Asp Leu Ile Val Gly Asn Tyr Ser Asp Gly
395 400 405
aac att gtt gct tct tta ttg gca ctg aag ttt gat gtt aca cag 1685
Asn Ile Val Ala Ser Leu Leu Ala Leu Lys Phe Asp Val Thr Gln
410 415 420
gtttgttaaa tgccaattaa ttgaacaagt tcctttctgt attttgattt ttcccgtgca 1745
atctaaaaac ttggttttca atgttttatg cag tgc act att gct cat gca ctt 1799
Cys Thr Ile Ala His Ala Leu
425
gag aag acg aaa tac ccg gat tca gac att aac tgg aaa caa ctt gag 1847
Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys Gln Leu Glu
430 435 440 445
gat aag tat cac ttc tcc tgt cag ttt act gct gat ctt att gct atg 1895
Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Ala Asp Leu Ile Ala Met
450 455 460
aac cat act gat ttt atc atc acc agc acc ttc caa gag att gct gga 1943
Asn His Thr Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu Ile Ala Gly
465 470 475
ag gtaatctatc tcccac ttta ctacatttgg ttcccttgta cgttgtaaat 1995
Ser
gttgccacct atttgagtcc ctaacaatgt ttccattcca tgattttcag c aag gac 2052
Lys Asp
480
act ctc ggc caa tac gag agt cac atc gct ttc act ctt cca ggg ctc 2100
Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro Gly Leu
485 490 495
tac cgc gtt gtt gat ggg atc gat gtt ttt gat ccc aaa ttc aat att 2148
Tyr Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe Asn Ile
500 505 510
gtc tcc cct ggt gct gat atg agc ata tac ttc cct tac acg gaa gag 2196
Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr Glu Glu
515 520 525
aag cga agg ttg aag aag ttc cac ccg gag att gaa gag ctt ctt tac 2244
Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu Leu Tyr
530 535 540
agc cct gtt gag aat aca gag cac tt gtaagatttt ggtcctagtt 2290
Ser Pro Val Glu Asn Thr Glu His Leu
545 550
ttacacccgt tttcaccgga ttttgtgttc agtgttaaca atgtcttaaa ttgttgtctc 2350
acag a tgt gta cta aaa gac cgc aac aag ccg gtt ctg ttt acc atg 2397
Cys Val Leu Lys Asp Arg Asn Lys Pro Val Leu Phe Thr Met
555 560 565
gca agg ttg gac cga gtg aag aac tta act ggg ctc gta gag ttc tat 2445
Ala Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Phe Tyr
570 575 580
gcc aag aac agc cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt 2493
Ala Lys Asn Ser Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly
585 590 595
gga gat agg aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg 2541
Gly Asp Arg Arg Lys Glu Ser Lys Asp Leu Glu Glu Lys Ala Glu Met
600 605 610 615
aag aag atg tat gaa ctt atc gaa aaa tac aag ttg aat gga caa ttg 2589
Lys Lys Met Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Leu
620 625 630
aga tgg ata tcg tcc cag atg aac cgt gtg aga aat ggt gaa ctc tat 2637
Arg Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr
635 640 645
cgt tat att tgc gac acc aag gga gca ttc gtt cag cct gct ata tac 2685
Arg Tyr Ile Cys Asp Thr Lys Gly Ala Phe Val Gln Pro Ala Ile Tyr
650 655 660
gag gct ttt ggc ttg act gtt gtt gag gca atg acc tgt gga cta cca 2733
Glu Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr Cys Gly Leu Pro
665 670 675
aca ttt gca aca tgt tac ggg ggc cct gct gag att ata gtt cac gga 2781
Thr Phe Ala Thr Cys Tyr Gly Gly Pro Ala Glu Ile Ile Val His Gly
680 685 690 695
aaa tcg ggg ttc aac atc gac cct tat aac ggt gat ttg gct gcc gag 2829
Lys Ser Gly Phe Asn Ile Asp Pro Tyr Asn Gly Asp Leu Ala Ala Glu
700 705 710
acc ctt gcc aat ttc ttc gag aaa tgc aaa gcg gat cca tct tat tgg 2877
Thr Leu Ala Asn Phe Phe Glu Lys Cys Lys Ala Asp Pro Ser Tyr Trp
715 720 725
gat gag atc tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca 2925
Asp Glu Ile Ser Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr
730 735 740
tgg cag att tac tcc gag aag cta ttg act ctc acc gga gtt tat ggc 2973
Trp Gln Ile Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly
745 750 755
ttt tcg aaa cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac 3021
Phe Ser Lys His Val Ala Tyr Gln Glu Gln Arg Gly Arg Lys Arg Tyr
760 765 770 775
att gaa atg ttg cat gca ctg atg tat aac aat cgg gtaaaacagt 3067
Ile Glu Met Leu His Ala Leu Met Tyr Asn Asn Arg
780 785
catgtctatt attattatta catcaaacat cattttactc tttcattgaa ctgaaaattt 3127
catatttctg aatgtttaat tgtgtgcttt acag gtc aag act gtt cct cta gct 3182
Val Lys Thr Val Pro Leu Ala
790
gtt gag taaagaaaaa gcaaaaagct gcttggaaac taaggaattt ccaagaatga 3238
Val Glu
795
ctgctgttga agttttattt ttgcttttat tatttttttt ttcaattggt aaatgtttaa 3298
tttcaacttt gggtttcccc ttggaatgtt tcgaactctg ggtttggtcc catt 3352
<210>9
<211>796
<212>PRT
＜213〉Lei Mengdeshi cotton (Gossypium raimondii)
<400>9
Met Ala Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Ala Thr
1 5 10 15
His Pro Gln Gln Ala Lys Ser Ile Leu Ser Arg Ile Glu Ser Leu Gly
20 25 30
Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp Lys Glu
35 40 45
Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu Arg Ser
50 55 60
Thr Gln Glu Ala Val Val Ser Ser Pro Leu Val Ala Leu Ala Ile Arg
65 70 75 80
Ser Ala Pro Gly Val Trp Glu Tyr Ile Ala Leu Glu Phe Gln Lys Leu
85 90 95
Phe Val Glu Glu Met Pro Val Ala Glu Tyr Leu Arg Leu Lys Glu Glu
100 105 110
Leu Val Asp Gly Ser Ser Asn Gly Glu Phe Met Leu Glu Leu Asp Phe
115 120 125
Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser Ile
130 135 140
Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu Phe
145 150 155 160
Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Glu Phe Leu Gln Ile Arg
165 170 175
Cys Gln Lys Gly Lys Gly Met Leu Leu Asn Asp Arg Ile Gln Asp Val
180 185 190
Asn Ser Leu Gln His Ala Leu Arg Lys Ala Glu Glu Tyr Leu Thr Pro
195 200 205
Leu Ser Ser Asp Thr Pro Tyr Ser Val Phe Glu Lys Arg Phe Leu Gly
210 215 220
Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Ala Glu His Val Leu Glu
225 230 235 240
Met Ile His Leu Leu Leu Asp Leu Leu Gln Ala Pro Asp Pro Val Ala
245 250 255
Leu Glu Ser Phe Leu Gly Arg Ile Pro Leu Val Ala Asn Val Val Ile
260 265 270
Met Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu Gly Tyr Pro
275 280 285
Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Ala Leu
290 295 300
Glu Glu Glu Leu Leu His Arg Phe Lys Leu Gln Gly Leu Asp Ile Thr
305 310 315 320
Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Ala Val Gly Thr
325 330 335
Thr Cys Gly Gln Arg Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp
340 345 350
Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp
355 360 365
Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp
370 375 380
Val Ala Ala Glu Ile Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile
385 390 395 400
Val Gly Asn Tyr Ser Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His
405 410 415
Lys Phe Asp Val Thr Gln Cys Ser Ile Ala His Ala Leu Glu Lys Thr
420 425 430
Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys Gln Leu Glu Asp Lys Tyr
435 440 445
His Phe Ser Cys Gln Phe Thr Ala Asp Leu Ile Ala Met Asn His Thr
450 455 460
Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu Ile Ala Gly Ser Lys Asp
465 470 475 480
Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro Gly Leu
485 490 495
Tyr Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe Asn Ile
500 505 510
Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr Glu Glu
515 520 525
Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu Leu Tyr
530 535 540
Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn
545 550 555 560
Lys Pro Ile Leu Phe Thr Met Ala Arg Leu Asp Arg Val Lys Asn Leu
565 570 575
Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu
580 585 590
Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp
595 600 605
Leu Glu Glu Lys Ala Glu Met Lys Lys Met Tyr Glu Leu Ile Glu Lys
610 615 620
Tyr Lys Leu Asn Gly Gln Phe Arg Trp Ile Ser Ser Gln Met Asn Arg
625 630 635 640
Val Arg Asn Gly Glu Leu Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Ala
645 650 655
Phe Val Gln Pro Ala Ile Tyr Glu Ala Phe Gly Leu Thr Val Val Glu
660 665 670
Ala Met Thr Cys Gly Leu Pro Thr Phe Ala Thr Cys Tyr Gly Gly Pro
675 680 685
Ala Glu Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Tyr
690 695 700
Asn Gly Asp Leu Ala Ala Glu Thr Leu Ala Asn Phe Phe Glu Lys Cys
705 710 715 720
Lys Ala Asp Pro Ser Tyr Trp Asp Glu Ile Ser Gln Gly Gly Leu Lys
725 730 735
Arg Ile Gln Glu Lys Tyr Thr Trp Gln Ile Tyr Ser Glu Lys Leu Leu
740 745 750
Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Ala Tyr Gln Glu
755 760 765
Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Leu His Ala Leu Met Tyr
770 775 780
Asn Asn Arg Val Lys Thr Ala Pro Leu Ala Val Glu
785 790 795
<210>10
<211>3354
<212>DNA
＜213〉Lei Mengdeshi cotton
<220>
＜221〉exon
<222>(32)..(111)
＜223〉exons 1
<220>
<221>misc_feature
<222>(32)..(34)
＜223〉initiator codon
<220>
＜221〉exon
<222>(197)..(314)
＜223〉exon 2
<220>
＜221〉exon
<222>(394)..(545)
＜223〉exon 3
<220>
＜221〉exon
<222>(625)..(817)
＜223〉exon 4
<220>
＜221〉exon
<222>(897)..(1328)
＜223〉exon 5
<220>
＜221〉exon
<222>(1399)..(1689)
＜223〉exon 6
<220>
＜221〉exon
<222>(1783)..(1949)
＜223〉exon 7
<220>
＜221〉exon
<222>(2050)..(2274)
＜223〉exon 8
<220>
＜221〉exon
<222>(2358)..(3060)
＜223〉exon 9
<220>
＜221〉exon
<222>(3169)..(3195)
＜223〉exons 10
<220>
<221>misc_feature
<222>(3196)..(3198)
＜223〉terminator codon
<400>10
cccttctgccatttcaggaa cctaaaaaac a atg gct tca atc agt gtt tgt 52
Met Ala Ser Ile Ser Val Cys
1 5
gag cgt ttg ggt gaa tct cta gct act cat cca cag cag gca aag tct 100
Glu Arg Leu Gly Glu Ser Leu Ala Thr His Pro Gln Gln Ala Lys Ser
10 15 20
atc ttg tca ag gtataatgta aataagtgct gccttcaaat ttctggtctt 151
Ile Leu Ser Arg
25
tctttgttat tttttattac attgtttata tatgtcatat tgcag g att gaa agc 206
Ile Glu Ser
30
ctc gga aag ggt att cat aag tct caa aag ctg ctc tcg gtt ctc gat 254
Leu Gly Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp
35 40 45
aaa gag gcc gga aat caa gca ctt gat ggg atg gtg gtg gag gte ctc 302
Lys Glu Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu
50 55 60
agg tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtttcatg 354
Arg Ser Thr Gln
65
atgagaatat tggactgata tgttaccgct atctaatag gaa gct gta gtg tcg 408
Glu Ala Val Val Ser
70
tct cca ttg gtt gcc ctt gcc att cgt tca gct cct gga gtt tgg gag 456
Ser Pro Leu Val Ala Leu Ala Ile Arg Ser Ala Pro Gly Val Trp Glu
75 80 85
tac att gct ctg gag ttc caa aag ctt ttt gtg gag gaa atg ccc gtt 504
Tyr Ile Ala Leu Glu Phe Gln Lys Leu Phe Val Glu Glu Met Pro Val
90 95 100
gct gag tat cta cgg ttg aag gaa gaa ctt gtt gat gga ag 545
Ala Glu Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser
105 110 115
gtaggtagtt ttctcttaca gctcctggtt tatattatgt ttgagtttat gttgaacaat 605
gctcaatgat gtggttcag c tcc aat ggc gag ttt atg ttg gaa ttg gac 655
Ser Asn Gly Glu Phe Met Leu Glu Leu Asp
120 125
ttt ggt gca ttc aat aat tct gtt cct cgt cca tct ctt tca aag tcc 703
Phe Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser
130 135 140
att ggt aat ggc atg gac ttc ctc aac cgc cac ctt tct gcc aag cta 751
Ile Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu
145 150 155
ttt caa gac aag gag aac ttg aac ttg ttg ctt gaa ttt ctc caa att 799
Phe Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Glu Phe Leu Gln Ile
160 165 170 175
cgc tgc cag aag gga aag gtaaaagttt cactttcact atttgttttt 847
Arg Cys Gln Lys Gly Lys
180
cctttctgtat acatcaaaa cttaagaatc gttcaatgtt atgccacag ggt atg ctg 905
Gly Met Leu
ttg aat gac aga atc caa gat gtg aat tcc ctc caa cat gca ttg agg 953
Leu Asn Asp Arg Ile Gln Asp Val Asn Ser Leu Gln His Ala Leu Arg
185 190 195 200
aag gcc gag gag tat ctg act cct cta tcg tcg gat acc ccg tac tca 1001
Lys Ala Glu Glu Tyr Leu Thr Pro Leu Ser Ser Asp Thr Pro Tyr Ser
205 210 215
gtt ttc gag aaa agg ttt ctg ggg att ggt ttg gag aag gga tgg ggc 1049
Val Phe Glu Lys Arg Phe Leu Gly Ile Gly Leu Glu Lys Gly Trp Gly
220 225 230
gat aat gct gag cat gtt ctt gag atg atc cat ctt cta ttg gat ctc 1097
Asp Asn Ala Glu His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu
235 240 245
ctt cag gca cct gat cct gtc gca ctt gaa agc ttc ctt gga aga atc 1145
Leu Gln Ala Pro Asp Pro Val Ala Leu Glu Ser Phe Leu Gly Arg Ile
250 255 260
cca ttg gtc gcc aat gtt gta atc atg act ccc cat gga tac ttt gcc 1193
Pro Leu Val Ala Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Ala
265 270 275 280
caa gac aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat 1241
Gln Asp Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr
285 290 295
atc tta gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt ttc 1289
Ile Leu Asp Gln Val Arg Ala Leu Glu Glu Glu Leu Leu His Arg Phe
300 305 310
aag ctg caa gga ctc gac att acc cca cgt atc cta gtc gtgagtatct 1338
Lys Leu Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val
315 320 325
gaatgttaat tcagtttcga tcattatgtt taatgcttaa tttggctgtt tgtgtaccag 1398
att act cgg ctc ctc cct gat gct gta gga aca act tgc gga cag cgt 1446
Ile Thr Arg Leu Leu Pro Asp Ala Val Gly Thr Thr Cys Gly Gln Arg
330 335 340
ctt gag aaa gtg tat gga acc aag tat tct gat att ctt cgg gta ccc 1494
Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp Ile Leu Arg Val Pro
345 350 355
ttc aga act gag aag gga att gta cgt cca tgg atc tca cga ttc aaa 1542
Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp Ile Ser Arg Phe Lys
360 365 370
gtc tgg cct tac ctg gag act tac act aag gat gtt gca gct gag atc 1590
Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp Val Ala Ala Glu Ile
375 380 385
acc aaa gag ttc cag ggc aag cct gat ctg att gtt ggg aac tac agt 1638
Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile Val Gly Asn Tyr Ser
390 395 400 405
gat gga aac att gtt gct tct tta ttg gca cat aag ttt gat gtt aca 1686
Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His Lys Phe Asp Val Thr
410 415 420
cag gtttgttaaatgccaattaa ttgaacaagt tccttgctgt attttgatgt 1739
Gln
tcctcgtgca atctaaaaac ttggttttca atgttttatg cag tgc tct att gct 1794
Cys Ser Ile Ala
425
cat gca ctc gag aag acg aaa tac ccg gat tca gac att aac tgg aaa 1842
His Ala Leu Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys
430 435 440
cag ctt gag gat aag tat cac ttc tcc tgt cag ttt act gct gat ctt 1890
Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Ala Asp Leu
445 450 455
att gct atg aac cat act gat ttt atc atc acc agc acc ttc caa gag 1938
Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu
460 465 470
att gct gga ag gtaatctatc tcccacttta ctacatttgg ttcccttgta 1989
Ile Ala Gly Ser
475
cgttgtaaat gttgccacct atttgagtcc ctaacaatgt tttcattcca tgattttcag 2049
c aag gac act ctc ggc caa tac gag agt cac att gct ttc act ctt cca 2098
Lys Asp Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro
480 485 490
ggt ctc tac cgc gtt gtt gat ggg atc gat gtt ttt gat ccc aaa ttc 2146
Gly Leu Tyr Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe
495 500 505 510
aat att gtc tcc cct ggt gct gat atg agc ata tac ttc cct tac acg 2194
Asn Ile Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr
515 520 525
gaa gag aag cgg agg ttg aag aag ttc cac ccg gag att gaa gag ctt 2242
Glu Glu Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu
530 535 540
ctt tac agc cct gtt gag aat aca gag cac tt gtgagatttt ggtcctattt 2294
Leu Tyr Ser Pro Val Glu Asn Thr Glu His Leu
545 550
ttacaccctt ttcactagattttgtgttca gtgttaacaa tgtcttaaat tgttgtctca 2354
cag a tgt gta cta aaa gac cgc aac aag ccg att ctg ttt acc atg gca 2403
Cys Val Leu Lys Asp Arg Asn Lys Pro Ile Leu Phe Thr Met Ala
555 560 565
agg ttg gac cga gtg aag aac tta act ggg ctc gta gag ttc tat gcc 2451
Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Phe Tyr Ala
570 575 580
aag aac agc cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt gga 2499
Lys Asn Ser Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly Gly
585 590 595 600
gat agg aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg aag 2547
Asp Arg Arg Lys Glu Ser Lys Asp Leu Glu Glu Lys Ala Glu Met Lys
605 610 615
aag atg tat gaa ctt atc gaa aaa tac aag ttg aat gga caa ttc aga 2595
Lys Met Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Phe Arg
620 625 630
tgg ata tcg tcc cag atg aac cgt gtg aga aat ggt gaa ctc tat cgt 2643
Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr Arg
635 640 645
tat att tgc gac acc aag gga gca ttc gtt cag cct gct ata tac gag 2691
Tyr Ile Cys Asp Thr Lys Gly Ala Phe Val Gln Pro Ala Ile Tyr Glu
650 655 660
gct ttt ggc ttg act gtt gtt gag gca atg acc tgt gga cta cca aca 2739
Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr Cys Gly Leu Pro Thr
665 670 675 680
ttt gca aca tgc tac ggg ggc cct gct gag att ata gtt cac gga aaa 2787
Phe Ala Thr Cys Tyr Gly Gly Pro Ala Glu Ile Ile Val His Gly Lys
685 690 695
tcg ggg ttc aac atc gat cct tat aac ggt gat ttg gctgcc gag acc 2835
Ser Gly Phe Asn Ile Asp Pro Tyr Asn Gly Asp Leu Ala Ala Glu Thr
700 705 710
ctt gcc aat ttc ttc gag aag tgc aaa gcg gat cca tct tat tgg gat 2883
Leu Ala Asn Phe Phe Glu Lys Cys Lys Ala Asp Pro Ser Tyr Trp Asp
715 720 725
gag atc tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca tgg 2931
Glu Ile Ser Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr Trp
730 735 740
cag att tac tcc gag aag cta ttg act ctc acc gga gtt tat ggc ttt 2979
Gln Ile Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly Phe
745 750 755 760
tcg aaa cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac att 3027
Ser Lys His Val Ala Tyr Gln Glu Gln Arg Gly Arg Lys Arg Tyr Ile
765 770 775
gaa atg ttg cat gca ttg atg tat aac aat cgg gtaaaacaat catgtctatt 3080
Glu Met Leu His Ala Leu Met Tyr Asn Asn Arg
780 785
attattatta catcaaacat cattttactc tttcattgaa ctgaaaattt catgtttctg 3140
aatgtttgtt taattgtgtg ttttacag gtc aag act gct cca cta gct gtt 3192
Val Lys Thr Ala Pro Leu Ala Val
790 795
gag taaagaaaaa gcaaaaggct gcttggaaac taaggaattt ccaagaatga 3245
Glu
ttactattga agttttattt ttgcttcttt tttttcaatt tggtaaatgt ttaatttcaa 3305
ctttggggtt tccccttgga atgttttgaa ctctgggttt ggtcccatt 3354
<210>11
<211>796
<212>PRT
＜213〉long calyx cotton (Gossypium longicalyx)
<400>11
Met Ala Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Ala Thr
1 5 10 15
His Pro Gln Gln Ala Lys Ser Ile Leu Ser Arg Ile Glu Ser Leu Gly
20 25 30
Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp Lys Glu
35 40 45
Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu Arg Ser
50 55 60
Thr Gln Glu Ala Val Val Ser Ser Pro Leu Val Ala Leu Ala Ile Arg
65 70 75 80
Ser Ala Pro Gly Val Trp Glu Tyr Ile Ala Val Glu Val Gln Lys Leu
85 90 95
Phe Val Glu Glu Met Pro Val Ala Glu Tyr Leu Arg Leu Lys Glu Glu
100 105 110
Leu Val Asp Gly Ser Ser Lys Gly Glu Phe Met Leu Glu Leu Asp Phe
115 120 125
Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser Ile
130 135 140
Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu Phe
145 150 155 160
Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Glu Phe Leu Gln Ile His
165 170 175
Cys Gln Lys Gly Lys Gly Met Leu Leu Asn Asp Arg Ile Gln Asp Val
180 185 190
Asn Ser Leu Gln His Ala Leu Arg Lys Ala Glu Glu Tyr Leu Thr Pro
195 200 205
Leu Thr Ser Asp Thr Pro Tyr Ser Val Phe Glu Lys Arg Phe Arg Glu
210 215 220
Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Ala Glu His Val Leu Glu
225 230 235 240
Met Ile His Leu Leu Leu Asp Leu Leu Gln Ala Pro Asp Pro Val Ala
245 250 255
Leu Glu Ser Phe Leu Gly Arg Ile Pro Leu Val Ala Asn Val Val Ile
260 265 270
Met Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu Gly Tyr Pro
275 280 285
Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Ala Leu
290 295 300
Glu Glu Glu Leu Leu His Arg Phe Lys Leu Gln Gly Leu Asp Ile Thr
305 310 315 320
Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Ala Val Gly Thr
325 330 335
Thr Cys Gly Gln Arg Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp
340 345 350
Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp
355 360 365
Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp
370 375 380
Val Ala Ala Glu Ile Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile
385 390 395 400
Val Gly Asn Tyr Ser Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His
405 410 415
Lys Phe Asp Val Thr Gln Cys Thr Ile Ala His Ala Leu Glu Lys Thr
420 425 430
Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys Gln Leu Glu Asp Lys Tyr
435 440 445
His Phe Ser Cys Gln Phe Thr Ala Asp Leu Ile Ala Met Asn His Thr
450 455 460
Asp Phe Val Ile Thr Ser Thr Phe Gln Glu Ile Ala Gly Ser Lys Asp
465 470 475 480
Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro Gly Leu
485 490 495
Tyr Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe Asn Ile
500 505 510
Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr Glu Glu
515 520 525
Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu Leu Tyr
530 535 540
Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn
545 550 555 560
Lys Pro Val Leu Phe Thr Met Ala Arg Leu Asp Arg Val Lys Asn Leu
565 570 575
Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu
580 585 590
Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp
595 600 605
Leu Glu Glu Lys Ala Glu Met Lys Lys Met Tyr Glu Leu Ile Glu Lys
610 615 620
Tyr Lys Leu Asn Gly Gln Phe Arg Trp Ile Ser Ser Gln Met Asn Arg
625 630 635 640
Val Arg Asn Gly Glu Leu Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Ala
645 650 655
Phe Val Gln Pro Ala Ile Tyr Glu Ala Phe Gly Leu Thr Val Val Glu
660 665 670
Ala Met Thr Cys Gly Leu Pro Thr Phe Ala Thr Cys Tyr Gly Gly Pro
675 680 685
Ala Glu Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Tyr
690 695 700
Asn Gly Asp Leu Ala Ala Glu Thr Leu Ala Asn Phe Phe Glu Lys Cys
705 710 715 720
Lys Ala Asp Pro Ser Tyr Trp Asp Glu Ile Ser Gln Gly Gly Leu Lys
725 730 735
Arg Ile Gln Glu Lys Tyr Thr Trp Gln Ile Tyr Ser Glu Lys Leu Leu
740 745 750
Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Ala Tyr Gln Glu
755 760 765
Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Leu His Ala Leu Met Tyr
770 775 780
Asn Asn Arg Val Lys Thr Val Pro Leu Ala Val Glu
785 790 795
<210>12
<211>3353
<212>DNA
＜213〉long calyx cotton
<220>
＜221〉exon
<222>(32)..(111)
＜223〉exon l
<220>
<221>misc_feature
<222>(32)..(34)
＜223〉ATG initiator codon
<220>
＜221〉exon
<222>(195)..(312)
＜223〉exon 2
<220>
＜221〉exon
<222>(392)..(543)
＜223〉exon 3
<220>
＜221〉exon
<222>(623)..(815)
＜223〉exon 4
<220>
＜221〉exon
<222>(891)..(1322)
＜223〉exon 5
<220>
＜221〉exon
<222>(1393)..(1683)
＜223〉exon 6
<220>
＜221〉exon
<222>(1777)..(1943)
＜223〉exon 7
<220>
＜221〉exon
<222>(2044)..(2268)
＜223〉exon 8
<220>
＜221〉exon
<222>(2353)..(3055)
＜223〉exon 9
<220>
＜221〉exon
<222>(3160)..(3186)
＜223〉exons 10
<220>
<221>misc_feature
<222>(3187)..(3189)
＜223〉terminator codon
<400>12
cccttctgcc atttcaggaa cctaaaaaac a atg gct tca atc agt gtc tgt 52
Met A1a Ser Ile Ser Val Cys
1 5
gag cgt ttg ggt gaa tct cta gct act cat cca cag cag gca aag tct 100
Glu Arg Leu Gly Glu Ser Leu Ala Thr His Pro Gln Gln Ala Lys Ser
10 15 20
atc ttg tca ag gtataatgta aataagtgct gccttcaaag ttctggtctt 151
Ile Leu Ser Arg
25
tctttgttat tttttattac attgtttata tgtcatattg cag g att gaa agc ctt 207
Ile Glu Ser Leu
30
gga aag ggt att cat aag tct caa aag ctg ctc tcg gtt ctc gat aaa 255
Gly Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp Lys
35 40 45
gag gcc gga aat caa gca ctt gat ggg atg gtg gtg gag gtc ctc agg 303
Glu Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu Arg
50 55 60
tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtttcatg 352
Ser Thr Gln
65
atgagaatat tggactgata tgttaccatt atctaatag gaa gct gta gtg tcg 406
Glu Ala Val Val Ser
70
tct cca ttg gtt gcc ctt gcc att cgt tca gct cct gga gtt tgg gag 454
Ser Pro Leu Val Ala Leu Ala Ile Arg Ser Ala Pro Gly Val Trp Glu
75 80 85
tac att gct gtg gag gtc caa aag ctt ttt gtg gag gaa atg ccc gtt 502
Tyr Ile Ala Val Glu Val Gln Lys Leu Phe Val Glu Glu Met Pro Val
90 95 100
gct gag tat cta cgg ttg aag gaa gaa ctt gtt gac gga ag 543
Ala Glu Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser
105 110 115
gtaggtagtt ttctcttaca gctcctggtt tatgttatgt ttgagtttat gttgaacaat 603
gctcaatgat gtggttcag c tcc aag ggc gag ttt atg ttg gaa ttg gac 653
Ser Lys Gly Glu Phe Met Leu Glu Leu Asp
120 125
ttt ggt gca ttc aat aat tct gtt cct cgt cca tct ctt tca aag tcc 701
Phe Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser
130 135 140
att ggt aat ggc atg gac ttc ctc aac cgc cac ctt tct gcc aag cta 749
Ile Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu
145 150 155
ttt caa gac aag gag aac ttg aac ttg ttg ctt gaa ttt ctc caa att 797
Phe Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Glu Phe Leu Gln Ile
160 165 170 175
cac tgc cag aag gga aag gtaaaagttt cactttcact atttgttttt 845
His Cys Gln Lys Gly Lys
180
cctttataca tcaaaactca agaatcgttc aatgttatgc cacag ggt atg ctg ttg 902
Gly Met Leu Leu
185
aat gac aga atc caa gat gtg aat tcc ctc caa cat gca ttg agg aag 950
Asn Asp Arg Ile Gln Asp Val Asn Ser Leu Gln His Ala Leu Arg Lys
190 195 200
gcc gag gag tat ctg act ccc cta acg tcg gat acc ccg tac tcg gtt 998
Ala Glu Glu Tyr Leu Thr Pro Leu Thr Ser Asp Thr Pro Tyr Ser Val
205 210 215
ttc gag aaa agg ttt cgg gag att ggt ttg gag aag gga tgg ggt gat 1046
Phe Glu Lys Arg Phe Arg Glu Ile Gly Leu Glu Lys Gly Trp Gly Asp
220 225 230
aat gct gag cat gtt ctt gag atg atc cat ctt cta ttg gat ctc ctt 1094
Asn Ala Glu His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu Leu
235 240 245
cag gca cct gat cct gtc gca ctt gaa agc ttc ctt gga aga atc cca 1142
Gln Ala Pro Asp Pro Val Ala Leu Glu Ser Phe Leu Gly Arg Ile Pro
250 255 260 265
ttg gtc gcc aat gtt gta atc atg act ccc cat gga tac ttt gct caa 1190
Leu Val Ala Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Ala Gln
270 275 280
gac aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat atc 1238
Asp Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr Ile
285 290 295
tta gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt ttc aag 1286
Leu Asp Gln Val Arg Ala Leu Glu Glu Glu Leu Leu His Arg Phe Lys
300 305 310
ctg caa gga ctc gac att acc cca cgt atc cta gtc gtgagtatct 1332
Leu Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val
315 320 325
gaatgttaat tcggtttcga tcattatgtt taatgcttaa tttggctgtt tatgtaccag 1392
att act cgg ctc ctc cct gat gct gta gga aca act tgc ggt cag cgt 1440
Ile Thr Arg Leu Leu Pro Asp Ala Val Gly Thr Thr Cys Gly Gln Arg
330 335 340
ctt gaa aaa gtg tat gga acc aag tat tct gat att ctt cgg gta ccc 1488
Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp Ile Leu Arg Val Pro
345 350 355
ttc aga act gag aag gga att gta cgt cca tgg atc tca cga ttc aaa 1536
Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp Ile Ser Arg Phe Lys
360 365 370
gtc tgg cct tac ctg gag act tac act aag gat gtt gca gct gag atc 1584
Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp Val Ala Ala Glu Ile
375 380 385
acc aaa gag ttc cag ggc aag cct gat ctg att gtt ggg aac tac agt 1632
Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile Val Gly Asn Tyr Ser
390 395 400 405
gat gga aac att gtt gct tct tta ttg gca cat aag ttt gat gtt aca 1680
Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His Lys Phe Asp Val Thr
410 415 420
cag gtttgttaaa tgccaattaa ttgaacaagt tccttgctgt attttgatgt 1733
Gln
ttcccgtgca atctaaaaac ttggttttca atgttttatg cag tgc act att gct 1788
Cys Thr Ile Ala
425
cat gca ctt gag aag acg aaa tac ccg gat tca gac att aac tgg aaa 1836
His Ala Leu Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys
430 435 440
cag ctt gag gat aag tat cac ttc tcc tgt cag ttt act gct gat ctt 1884
Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Ala Asp Leu
445 450 455
att gct atg aac cat act gat ttt gtc atc acc agc acc ttc caa gag 1932
Ile Ala Met Asn His Thr Asp Phe Val Ile Thr Ser Thr Phe Gln Glu
460 465 470
att gct gga ag gtaatctatc tcccacttta ctaaatttgg ttcccttgta 1983
Ile Ala Gly Ser
475
cgttgtaaat gttgccacct atttgagtcc ctaacaatgt ttccattcca tgattttcag 2043
c aag gac act ctc ggc caa tac gag agt cac att gct ttc act ctt cca 2092
Lys Asp Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro
480 485 490
ggg ctc tac cgc gtt gtt gat ggg atc gat gtt ttt gat ccc aaa ttc 2140
Gly Leu Tyr Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe
495 500 505 510
aat att gtc tcc cct ggt gct gat atg agc ata tac ttc cct tac acg 2188
Asn Ile Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr
515 520 525
gaa gag aag cga agg ttg aag aag ttc cac ccg gag att gaa gag ctt 2236
Glu Glu Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu
530 535 540
ctt tac agc cct gtt gag aat aca gag cac tt gtgggatttt ggtcctattt 2288
Leu Tyr Ser Pro Val Glu Asn Thr Glu His Leu
545 550
ttacaccctt tttcactaga ttttgtgttc agtgttaaca atgtcttgaa ttgttgtctc 2348
acag a tgt gta cta aaa gac cgc aac aag ccg gtt ctg ttt acc atg 2395
Cys Val Leu Lys Asp Arg Asn Lys Pro Val Leu Phe Thr Met
555 560 565
gca agg ttg gac cga gtg aag aac tta act ggg ctc gta gag ttc tat 2443
Ala Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Phe Tyr
570 575 580
gcc aag aac agc cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt 2491
Ala Lys Asn Ser Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly
585 590 595
gga gat agg aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg 2539
Gly Asp Arg Arg Lys Glu Ser Lys Asp Leu Glu Glu Lys Ala Glu Met
600 605 610 615
aag aag atg tat gaa ctt atc gaa aaa tac aag ttg aat gga caa ttc 2587
Lys Lys Met Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Phe
620 625 630
aga tgg ata tcg tcc cag atg aac cgt gtg aga aat ggt gaa ctc tat 2635
Arg Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr
635 640 645
cgt tat att tgc gac acc aag gga gca ttc gtt cag cct gct ata tac 2683
Arg Tyr Ile Cys Asp Thr Lys Gly Ala Phe Val Gln Pro Ala Ile Tyr
650 655 660
gag gct ttt ggc ttg act gtt gtt gag gca atg acc tgt gga cta cca 2731
Glu Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr Cys Gly Leu Pro
665 670 675
aca ttt gca aca tgt tac ggg ggc cct gct gag att ata gtt cac gga 2779
Thr Phe Ala Thr Cys Tyr Gly Gly Pro Ala Glu Ile Ile Val His Gly
680 685 690 695
aaa tcg ggg ttc aac atc gac cct tat aac ggt gat ttg gct gcc gag 2827
Lys Ser Gly Phe Asn Ile Asp Pro Tyr Asn Gly Asp Leu Ala Ala Glu
700 705 710
acc ctc gcc aat ttc ttc gag aaa tgc aaa gcg gat cca tct tat tgg 2875
Thr Leu Ala Asn Phe Phe Glu Lys Cys Lys Ala Asp Pro Ser Tyr Trp
715 720 725
gat gag atc tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca 2923
Asp Glu Ile Ser Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr
730 735 740
tgg cag att tac tcc gag aag cta ttg act ctc acc gga gtt tat ggc 2971
Trp Gln Ile Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly
745 750 755
ttt tcg aaa cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac 3019
Phe Ser Lys His Val Ala Tyr Gln Glu Gln Arg Gly Arg Lys Arg Tyr
760 765 770 775
att gaa atg ttg cat gca ttg atg tat aac aat cgg gtaaaacaat 3065
Ile Glu Met Leu His Ala Leu Met Tyr Asn Asn Arg
780 785
catgtctatt attattatta catcaaacat cattttactc tttcattgaa ctgaaaattt 3125
catgtttctg aatgtttaat tgtgtgcttt acag gtc aag act gtt cct cta gct 3180
Val Lys Thr Val Pro Leu Ala
790
gtt gag taaagaaaca gcaaaaagct gcttggaaac caaggaattt ccaagaatga 3236
Val Glu
795
ttgctattga agttttattt ttgcttttct tttttttttt ttttttcaat ttggtaaatg 3296
tttaatttca actttggggt ttccccttgg aatgttttga actctgggtt tggtccc 3353
<210>13
<211>796
<212>PRT
＜213〉long calyx cotton
<400>13
Met Ala Ser Ile Ser Val Cys Glu Arg Leu Gly Glu Ser Leu Ala Thr
1 5 10 15
His Pro Gln Gln Ala Lys Ser Ile Leu Ser Arg Ile Glu Ser Leu Gly
20 25 30
Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp Lys Glu
35 40 45
Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu Arg Ser
50 55 60
Thr Gln Glu Ala Val Val Ser Ser Pro Leu Val Ala Leu Ala Ile Arg
65 70 75 80
Scr Ala Pro Gly Val Trp Glu Tyr Ile Ala Val Glu Val Gln Lys Leu
85 90 95
Phe Val Glu Glu Met Pro Val Ala Glu Tyr Leu Arg Leu Lys Glu Glu
100 105 110
Leu Val Asp Gly Ser Ser Asn Gly Glu Phe Met Leu Glu Leu Asp Phe
115 120 125
Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser Ile
130 135 140
Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu Phe
145 150 155 160
Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Glu Phe Leu Gln Ile His
165 170 175
Cys Gln Lys Gly Lys Gly Met Leu Leu Asn Asp Arg Ile Gln Asp Val
180 185 190
Asn Ser Leu Gln His Ala Leu Arg Lys Ala Glu Glu Tyr Leu Thr Pro
195 200 205
Leu Ser Ser Asp Thr Pro Tyr Ser Val Phe Glu Lys Arg Phe Leu Gly
210 215 220
Ile Gly Leu Glu Lys Gly Trp Gly Asp Asn Ala Glu His Val Leu Glu
225 230 235 240
Met Ile His Leu Leu Leu Asp Leu Leu Gln Ala Pro Asp Pro Val Ala
245 250 255
Leu Glu Ser Phe Leu Gly Arg Ile Pro Leu Val Ala Asn Val Val Ile
260 265 270
Met Thr Pro His Gly Tyr Phe Ala Gln Asp Asn Val Leu Gly Tyr Pro
275 280 285
Asp Thr Gly Gly Gln Val Val Tyr Ile Leu Asp Gln Val Arg Ala Leu
290 295 300
Glu Glu Glu Leu Leu His Arg Ser Lys Leu Gln Gly Leu Asp Ile Thr
305 310 315 320
Pro Arg Ile Leu Val Ile Thr Arg Leu Leu Pro Asp Ala Val Gly Thr
325 330 335
Thr Cys Gly Gln Arg Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp
340 345 350
Ile Leu Arg Val Pro Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp
355 360 365
Ile Ser Arg Phe Lys Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp
370 375 380
Val Ala Ala Glu Ile Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile
385 390 395 400
Val Gly Asn Tyr Ser Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His
405 410 415
Lys Phe Asp Val Thr Gln Cys Thr Ile Ala His Ala Leu Glu Lys Thr
420 425 430
Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys Gln Leu Glu Asp Lys Tyr
435 440 445
His Phe Ser Cys Gln Phe Thr Ala Asp Leu Ile Ala Met Asn His Thr
450 455 460
Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu Ile Ala Gly Ser Lys Asp
465 470 475 480
Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro Gly Leu
485 490 495
Cys Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe Asn Ile
500 505 510
Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr Glu Glu
515 520 525
Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu Leu Tyr
530 535 540
Ser Pro Val Glu Asn Thr Glu His Leu Cys Val Leu Lys Asp Arg Asn
545 550 555 560
Lys Pro Ile Leu Phe Thr Met Ala Arg Leu Asp Arg Val Lys Asn Leu
565 570 575
Thr Gly Leu Val Glu Phe Tyr Ala Lys Asn Ser Arg Leu Arg Glu Leu
580 585 590
Val Asn Leu Val Val Val Gly Gly Asp Arg Arg Lys Glu Ser Lys Asp
595 600 605
Leu Glu Glu Lys Ala Glu Met Lys Lys Met Tyr Glu Leu Ile Glu Lys
610 615 620
Tyr Lys Leu Asn Gly Gln Phe Arg Trp Ile Ser Ser Gln Met Asn Arg
625 630 635 640
Val Arg Asn Gly Glu Leu Tyr Arg Tyr Ile Cys Asp Thr Lys Gly Ala
645 650 655
Phe Val Gln Pro Pro Ile Tyr Glu Ala Phe Gly Leu Thr Va1 Val Glu
660 665 670
Ala Met Thr Cys Gly Leu Pro Thr Phe Ala Thr Cys Tyr Gly Gly Pro
675 680 685
Ala Glu Ile Ile Val His Gly Lys Ser Gly Phe Asn Ile Asp Pro Cys
690 695 700
Asn Gly Asp Leu Ala Ala Glu Thr Leu Ala Asn Phe Phe Glu Lys Cys
705 710 715 720
Lys Ala Asp Pro Ser Tyr Trp Asp Glu Ile Ser Gln Gly Gly Leu Lys
725 730 735
Arg Ile Gln Glu Lys Tyr Thr Trp Gln Ile Tyr Ser Glu Lys Leu Leu
740 745 750
Thr Leu Thr Gly Val Tyr Gly Phe Ser Lys His Val Ala Tyr Gln Glu
755 760 765
Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Leu His Ala Trp Met Tyr
770 775 780
Asn Asn Arg Val Lys Thr Val Pro Leu Ala Val Glu
785 790 795
<210>14
<211>3353
<212>DNA
＜213〉long calyx cotton
<220>
＜221〉exon
<222>(32)..(111)
＜223〉exons 1
<220>
<221>misc_feature
<222>(32)..(34)
＜223〉initiator codon
<220>
＜221〉exon
<222>(197)..(314)
＜223〉exon 2
<220>
＜221〉exon
<222>(394)..(545)
＜223〉exon 3
<220>
＜221〉exon
<222>(625)..(817)
＜223〉exon 4
<220>
＜221〉exon
<222>(897)..(1328)
＜223〉exon 5
<220>
＜221〉exon
<222>(1399)..(1689)
＜223〉exon 6
<220>
＜221〉exon
<222>(1783)..(1949)
＜223〉exon 7
<220>
＜221〉exon
<222>(2050)..(2274)
＜223〉exon 8
<220>
＜221〉exon
<222>(2358)..(3060)
＜223〉exon 9
<220>
＜221〉exon
<222>(3169)..(3195)
＜223〉exons 10
<220>
<221>misc_feature
<222>(3196)..(3198)
＜223〉terminator codon
<400>14
cccttctgcc atttcaggaa cccaaaaaac a atg gct tca atc agt gtt tgt 52
Met Ala Ser Ile Ser Val Cys
1 5
gag cgt ttg ggt gaa tct cta gct act cac cca cag cag gca aag tct 100
Glu Arg Leu Gly Glu Ser Leu Ala Thr His Pro Gln Gln Ala Lys Ser
10 15 20
atc ttg tca ag gtataatgta aataagtgct gccttcaaat ttctggtctt 151
Ile Leu Ser Arg
25
tcttcgttat tttttattac attgtttata tatgtcatat tgcag g att gaa agc 206
Ile Glu Ser
30
ctc gga aag ggt att cat aag tct caa aag ctg ctc tcg gtt ctc gat 254
Leu Gly Lys Gly Ile His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp
35 40 45
aaa gag gcc gga aat caa gca ctt gat ggg atg gtg gtg gag gtc ctc 302
Lys Glu Ala Gly Asn Gln Ala Leu Asp Gly Met Val Val Glu Val Leu
50 55 60
agg tcc act cag gtcacaataa aacaaaccct ttctttggtt atgtttcatg 354
Arg Ser Thr Gln
65
atgagaatat tggactgata tgttaccgtt atctaatag gaa gct gta gtg tcg 408
Glu Ala Val Val Ser
70
tct cca ttg gtt gcc ctt gcc att cgt tca gct cct gga gtt tgg gag 456
Ser Pro Leu Val Ala Leu Ala Ile Arg Ser Ala Pro Gly Val Trp Glu
75 80 85
tac att gct gtg gag gtc caa aag ctt ttt gtg gag gaa atg ccc gtt 504
Tyr Ile Ala Val Glu Val Gln Lys Leu Phe Val Glu Glu Met Pro Val
90 95 100
gct gag tat cta cgg ttg aag gaa gaa ctt gtt gat gga ag 545
Ala Glu Tyr Leu Arg Leu Lys Glu Glu Leu Val Asp Gly Ser
105 110 115
gtaggtagtt ttctcttaca gctcctggtt tatattatgt ttgagtttat gttgaacaat 605
gctcaatgat gtggttcag c tcc aat ggc gag ttt atg ttg gaa ttg gac 655
Ser Asn Gly Glu Phe Met Leu Glu Leu Asp
120 125
ttt ggt gca ttc aat aat tct gtt ccc cgt cca tct ctt tca aag tcc 703
Phe Gly Ala Phe Asn Asn Ser Val Pro Arg Pro Ser Leu Ser Lys Ser
130 135 140
att ggt aat ggc atg gac ttc ctc aac cgc cac ctt tct gcc aag cta 751
Ile Gly Asn Gly Met Asp Phe Leu Asn Arg His Leu Ser Ala Lys Leu
145 150 155
ttt caa gac aag gag aac ttg aac ttg ttg ctt gaa ttt ctc caa att 799
Phe Gln Asp Lys Glu Asn Leu Asn Leu Leu Leu Glu Phe Leu Gln Ile
160 165 170 175
cac tgc cag aag gga aag gtaaaagttt cactttcact atttgttttt 847
HisCys Gln Lys Gly Lys
180
cctttctgta tacatcaaaa cttaagaatc gttcaatgtt atgccacag ggt atg ctg 905
Gly Met Leu
ttg aat gac aga atc caa gat gtg aat tcc ctc caa cat gca tta agg 953
Leu Asn Asp Arg Ile Gln Asp Val Asn Ser Leu Gln His Ala Leu Arg
185 190 195 200
aag gcc gag gag tat ctg act cct cta tcc tcg gat acc ccg tac tca 1001
Lys Ala Glu Glu Tyr Leu Thr Pro Leu Ser Ser Asp Thr Pro Tyr Ser
205 210 215
gtt ttc gag aaa agg ttt ctg ggg att ggt ttg gag aag gga tgg ggt 1049
Val Phe Glu Lys Arg Phe Leu Gly Ile Gly Leu Glu Lys Gly Trp Gly
220 225 230
gat aat gct gag cat gtt ctt gag atg atc cat ctt cta ttg gat ctc 1097
Asp Asn Ala Glu His Val Leu Glu Met Ile His Leu Leu Leu Asp Leu
235 240 245
ctt cag gca cct gat cct gtc gca ctt gaa agc ttc ctt gga aga atc 1145
Leu Gln Ala Pro Asp Pro Val Ala Leu Glu Ser Phe Leu Gly Arg Ile
250 255 260
cca ttg gtc gcc aat gtt gta atc atg act ccc cat gga tac ttt gcc 1193
Pro Leu Val Ala Asn Val Val Ile Met Thr Pro His Gly Tyr Phe Ala
265 270 275 280
caa gac aat gtt ttg gga tac cct gac aca ggt ggt cag gtt gtt tat 1241
Gln Asp Asn Val Leu Gly Tyr Pro Asp Thr Gly Gly Gln Val Val Tyr
285 290 295
atc tta gat caa gtt cgt gcc ttg gag gaa gag ttg ctt cat cgt tcc 1289
Ile Leu Asp Gln Val Arg Ala Leu Glu Glu Glu Leu Leu His Arg Ser
300 305 310
aag ctg caa gga ctc gac att acc cca cgt atc cta gtc gtgagtatct 1338
Lys Leu Gln Gly Leu Asp Ile Thr Pro Arg Ile Leu Val
315 320 325
gaatgttaat tcagtttcga tcattatgtt taatgcttaa tttggctgtt tgtgtaccag 1398
att act cgg ctc ctc cct gat gct gta gga aca act tgc ggt cag cgt 1446
Ile Thr Arg Leu Leu Pro Asp Ala Val Gly Thr Thr Cys Gly Gln Arg
330 335 340
ctt gag aaa gtg tat gga acc aag tat tct gat att ctt cgg gta ccc 1494
Leu Glu Lys Val Tyr Gly Thr Lys Tyr Ser Asp Ile Leu Arg Val Pro
345 350 355
ttc aga act gag aag gga att gta cgt cca tgg atc tca cga ttc aaa 1542
Phe Arg Thr Glu Lys Gly Ile Val Arg Pro Trp Ile Ser Arg Phe Lys
360 365 370
gtc tgg cct tac ctg gag act tac act aag gat gtt gca gct gag atc 1590
Val Trp Pro Tyr Leu Glu Thr Tyr Thr Lys Asp Val Ala Ala Glu Ile
375 380 385
acc aaa gag ttc cag ggc aag cct gat ctg att gtt ggg aac tac agt 1638
Thr Lys Glu Phe Gln Gly Lys Pro Asp Leu Ile Val Gly Asn Tyr Ser
390 395 400 405
gat gga aac att gtt gct tct tta ttg gca cat aag ttt gat gtt aca 1686
Asp Gly Asn Ile Val Ala Ser Leu Leu Ala His Lys Phe Asp Val Thr
410 415 420
cag gtttgttaaa taccaattaa ttgaacaagt tccttgctgt attttgatgt 1739
Gln
tccccgtgca atctaaaaac ttggttttca atgttttatg cag tgc act att gct 1794
Cys Thr Ile Ala
425
cat gca ctc gag aag acg aaa tac ccg gat tca gac att aac tgg aaa 1842
His Ala Leu Glu Lys Thr Lys Tyr Pro Asp Ser Asp Ile Asn Trp Lys
430 435 440
cag ctt gag gat aag tat cac ttc tcc tgt cag ttt act gct gat ctt 1890
Gln Leu Glu Asp Lys Tyr His Phe Ser Cys Gln Phe Thr Ala Asp Leu
445 450 455
att gct atg aac cat act gat ttt atc atc acc agc acc ttc caa gag 1938
Ile Ala Met Asn His Thr Asp Phe Ile Ile Thr Ser Thr Phe Gln Glu
460 465 470
att gct gga ag gtaatctatc tcccacttta ctacatttgg ttcccttgta 1989
Ile Ala Gly Ser
475
cgttgtaaat gttgccacct atttgagtcc ctaacaatgt tttcattcca tgattttcag 2049
c aag gac act ctc ggc caa tac gag agt cac att gct ttc act ctt cca 2098
Lys Asp Thr Leu Gly Gln Tyr Glu Ser His Ile Ala Phe Thr Leu Pro
480 485 490
ggg ctc tgc cgc gtt gtt gat ggg atc gat gtt ttt gat ccc aaa ttc 2146
Gly Leu Cys Arg Val Val Asp Gly Ile Asp Val Phe Asp Pro Lys Phe
495 500 505 510
aat att gtc tcc cct ggt gct gat atg agc ata tac ttc cct tac acg 2194
Asn Ile Val Ser Pro Gly Ala Asp Met Ser Ile Tyr Phe Pro Tyr Thr
515 520 525
gaa gag aag cgg agg ttg aag aag ttc cac ccg gag att gaa gag ctt 2242
Glu Glu Lys Arg Arg Leu Lys Lys Phe His Pro Glu Ile Glu Glu Leu
530 535 540
ctt tac agc cct gtt gag aat aca gag cac tt gtgagatttt ggttctattt 2294
Leu Tyr Ser Pro Val Glu Asn Thr Glu His Leu
545 550
ttacaccctt ttcactagat tttgtgttca gtgttaacaa tgtcttaaat tgttgtctca 2354
cag a tgt gta cta aaa gac cgc aac aag ccg att ctg ttt acc atg gca 2403
Cys Val Leu Lys Asp Arg Asn Lys Pro Ile Leu Phe Thr Met Ala
555 560 565
agg ttg gac cga gtg aag aac tta act ggg ctc gta gag ttc tat gcc 2451
Arg Leu Asp Arg Val Lys Asn Leu Thr Gly Leu Val Glu Phe Tyr Ala
570 575 580
aag aac agc cgg ctg agg gaa ctg gtt aac ttg gtt gta gta ggt gga 2499
Lys Asn Ser Arg Leu Arg Glu Leu Val Asn Leu Val Val Val Gly Gly
585 590 595 600
gat agg aga aag gaa tcc aag gac tta gaa gaa aag gct gaa atg aag 2547
Asp Arg Arg Lys Glu Ser Lys Asp Leu Glu Glu Lys Ala Glu Met Lys
605 610 615
aag atg tat gaa ctt atc gaa aaa tac aag ttg aat gga caa ttc aga 2595
Lys Met Tyr Glu Leu Ile Glu Lys Tyr Lys Leu Asn Gly Gln Phe Arg
620 625 630
tgg ata tcg tcc cag atg aac cga gtg aga aat ggt gaa ctc tat cgt 2643
Trp Ile Ser Ser Gln Met Asn Arg Val Arg Asn Gly Glu Leu Tyr Arg
635 640 645
tat at ttgc gac acc aag gga gca ttc gtt cag cct cct ata tac gag 2691
Tyr Ile Cys Asp Thr Lys Gly Ala Phe Val Gln Pro Pro Ile Tyr Glu
650 655 660
gct ttt ggc ttg act gtt gtt gag gca atg acc tgt gga cta cca aca 2739
Ala Phe Gly Leu Thr Val Val Glu Ala Met Thr Cys Gly Leu Pro Thr
665 670 675 680
ttt gca aca tgt tac ggg ggc cct gct gag att ata gtt cac gga aaa 2787
Phe Ala Thr Cys Tyr Gly Gly Pro Ala Glu Ile Ile Val His Gly Lys
685 690 695
tcg ggg ttc aac atc gat cct tgt aac ggt gat ttg gct gcc gag acc 2835
Ser Gly Phe Asn Ile Asp Pro Cys Asn Gly Asp Leu Ala Ala Glu Thr
700 705 710
ctt gcc aat ttc ttc gag aag tgc aaa gcg gat cca tct tat tgg gat 2883
Leu Ala Asn Phe Phe Glu Lys Cys Lys Ala Asp Pro Ser Tyr Trp Asp
715 720 725
gag atc tcc cag gga ggg ttg aaa cgc ata cag gag aag tat aca tgg 2931
Glu Ile Ser Gln Gly Gly Leu Lys Arg Ile Gln Glu Lys Tyr Thr Trp
730 735 740
cag att tac tcc gag aag cta ttg act ctc acc gga gtt tat ggc ttt 2979
Gln Ile Tyr Ser Glu Lys Leu Leu Thr Leu Thr Gly Val Tyr Gly Phe
745 750 755 760
tcg aaa cat gta gct tac cag gag caa cgt ggg cgc aag cgt tac att 3027
Ser Lys His Val Ala Tyr Gln Glu Gln Arg Gly Arg Lys Arg Tyr Ile
765 770 775
gaa atg ttg cat gca tgg atg tat aac aat cgg gtaaaacaat catgtctatt 3080
Glu Met Leu His Ala Trp Met Tyr Asn Asn Arg
780 785
attattatta catcaaacat cattttactc tttcattgaa ctgaaatttt catgtttctg 3140
aatgtttgtt taattgtgtg ctttacag gtc aag act gtt cca cta gct gtt 3192
Val Lys Thr Val Pro Leu Ala Val
790 795
gag taaagaaaaa gcaaaaggct gcttggaaac caaggaattt ccaagaatga 3245
Glu
ttgctattga agttttattt ttgcttcttt ttttcaattt cgtaaatgtt taatttcaac 3305
tttggggttt ccccttggaa tgttttgaac tctgggtttg gtcccatt 3353
<210>15
<211>26
<212>PRT
＜213〉artificial sequence
<220>
＜223〉C of SusC end consensus sequence
<400>15
Ala Tyr Gln Glu Gln Arg Gly Arg Lys Arg Tyr Ile Glu Met Leu His
1 5 10 15
Ala Trp Met Tyr Asn Asn Arg Val Lys Thr
20 25
<210>16
<211>24
<212>PRT
＜213〉artificial sequence
<220>
＜223〉N of SusC end consensus sequence
<400>16
His Lys Ser Gln Lys Leu Leu Ser Val Leu Asp Lys Glu Ala Gly Asn
1 5 10 15
Gln Ala Leu Asp Gly Met Val Val
20
<210>17
<211>22
<212>DNA
＜213〉artificial sequence
<220>
＜223〉SusC specific PCR primer 1
<400>17
cccttctgccatttcaggaa cc 22
<210>18
<211>22
<212>DNA
＜213〉artificial sequence
<220>
＜223〉SusC specific PCR primer 2
<400>18
aatgggacca aacccagagt tc 22

Claims (34)

1. the isolating sucrose synthase albumen that has following aminoacid sequence, described aminoacid sequence comprise and are selected from following aminoacid sequence:
A. the aminoacid sequence that has at least 80% sequence homology with SEQ ID Nos.:2, any aminoacid sequence of 3,5,7,9,11 or 13;
B. comprise SEQ ID Nos.:2, any aminoacid sequence of aminoacid sequence of 3,5,7,9,11 or 13;
C. be positioned at the aminoacid sequence of described proteic aminoterminal part, described aminoacid sequence comprises the sequence identity with the aminoacid sequence about at least 60% of SEQ ID No.16; Or
D. be positioned at the aminoacid sequence of described proteic carboxyl terminal part, described aminoacid sequence comprises the sequence identity with the aminoacid sequence about at least 60% of SEQ ID No.15.
2. the isolating sucrose synthase albumen of claim 1, wherein said albumen comprises hydrophobicity N terminal sequence.
3. the isolating sucrose synthase albumen of claim 1 or claim 2, wherein said albumen also comprises any in the following amino acid sequences:
394 the aminoacid sequence of a.SEQ ID No.:3 from amino acid 383 to amino acid;
329 the aminoacid sequence of b.SEQ ID No.:3 from amino acid 270 to amino acid;
737 the aminoacid sequence of c.SEQ ID No.:3 from amino acid 549 to amino acid;
545 the aminoacid sequence of d.SEQ ID No.:3 from amino acid/11 to amino acid; Or
794 the aminoacid sequence of e.SEQ ID No.:3 from amino acid/11 8 to amino acid.
4. each isolating proteic antibody of identification claim 1 to 3.
5. each proteic isolated DNA molecule of sucrose synthase or nucleic acid of coding claim 1 to 3.
6. the isolated DNA molecule of claim 5 or nucleic acid, it comprises SEQ ID Nos.:1,4,6,8,10,12 or 14 each nucleotide sequence.
7. the expression cassette that comprises the dna molecular of following effective connection:
A) plant can be expressed promotor;
B) DNA of the sucrose synthase of each of coding claim 1 to 3 or the separated DNA of claim 6; Randomly
C) Transcription Termination and polyadenylation region.
8. the expression cassette of claim 7, wherein plant can be expressed promotor and preferentially controlled in fiber production fibre of plant cell and transcribe.
9. the expression cassette that comprises the dna molecular of following effective connection:
A) plant can be expressed promotor
Produce the DNA of RNA molecule when b) transcribing, wherein said RNA molecule comprises and is selected from following nucleotide sequence:
I. the nucleotide sequence of at least 19 continuous nucleotides, the wherein nucleotide sequence of this nucleotide sequence and endogenous sucrose synthase C isotype encoding gene or have about at least 94% sequence identity with the complementary sequence of the nucleotide sequence of described endogenous sucrose synthase C isotype encoding gene;
Ii. the nucleotide sequence of at least 19 continuous nucleotides, the nucleotide sequence of wherein said nucleotide sequence and claim 5 or claim 6 has about at least 94% sequence identity; Randomly
Iii. Transcription Termination and polyadenylation region.
10. the expression cassette that comprises the dna molecular of following effective connection:
A) plant can be expressed promotor, and the preferred preferentially plant that control is transcribed in fibrocyte can be expressed promotor;
Produce the DNA of the double stranded rna molecule of the endogenous SusC genetic expression that can reduce the fiber production plant when b) transcribing, wherein this RNA molecule comprises the first and second RNA zones, wherein
I. a RNA zone comprises the nucleotide sequence of at least 19 continuous nucleotides, the nucleotide sequence of this nucleotide sequence and endogenous SusC gene or have about at least 94% sequence identity with the nucleotide sequence of claim 5 or claim 6;
Ii. the 2nd RNA zone comprises these at least 19 the continuous nucleotide complementary nucleotide sequences with a RNA zone;
Iii. the first and second RNA zones can base pairing, thereby forms double stranded rna molecule between these at least 19 continuous nucleotides in first and second zones; With
C) be included in the 3 ' stub area that has the Transcription Termination and the polyadenylation signal of function in the cell of plant.
11. the expression cassette of claim 9 or claim 10, wherein plant can be expressed promotor and preferentially controlled in fiber production fibre of plant cell and transcribe.
12. comprise the expression cassette in the DNA zone of following effective connection:
A. plant can be expressed promotor;
B. importing vegetable cell and in vegetable cell, transcribing post-treatment and become the DNA zone of miRNA, wherein this miRNA can discern plant endogenous SusC encoding gene mRNA and instruct cutting to this mRNA; With
C. randomly, participate in 3 ' DNA zone of Transcription Termination and polyadenylation.
13. the expression cassette of claim 12, wherein said DNA zone comprises the encoding gene with SusC, the SusC gene that for example comprises SEQ ID Nos.:1, arbitrary nucleotide sequence of 4,6,8,10,12 or 14, the nucleotide sequence complementary nucleotide sequence basically of at least 21 continuous nucleotides, condition is to allow one or more following mispairing:
A) in the 5 ' terminal nucleotide of miRNA and the mispairing between the corresponding nucleotide sequence in the RNA molecule;
B) in any Nucleotide of position 1 to the position 9 of miRNA and the mispairing between the corresponding nucleotide sequence in the RNA molecule; Or
C) three in any Nucleotide of position 12 to the position 21 of miRNA and the mispairing between the corresponding nucleotide sequence in the RNA molecule, and condition is not exist to surpass two continuous mispairing.
14. comprise the vegetable cell that the allos plant that effectively is connected with dna molecular can be expressed promotor, described dna molecular is selected from following dna molecular:
A) DNA of the sucrose synthase of each of coding claim 1 to 3;
B) DNA of claim 6;
Produce the DNA of RNA molecule when c) transcribing, described RNA molecule comprises the nucleotide sequence of at least 19 continuous nucleotides, and the complementary sequence of the nucleotide sequence of the gene of described nucleotide sequence and the endogenous sucrose synthase C isotype of coding or the nucleotide sequence of described endogenous sucrose synthase C isotype encoding gene has about at least 94% sequence identity;
Produce the DNA of RNA molecule when d) transcribing, described RNA molecule comprises the nucleotide sequence of at least 19 continuous nucleotides, and the nucleotide sequence of described nucleotide sequence and claim 5 or claim 6 has about at least 94% sequence identity;
Produce the DNA of double stranded rna molecule when e) transcribing, described double stranded rna molecule can reduce the endogenous SusC genetic expression of fiber production plant, and this RNA molecule comprises the first and second RNA zones, wherein
I. a RNA zone comprises the nucleotide sequence of at least 19 continuous nucleotides, and the nucleotide sequence of described nucleotide sequence and endogenous SusC gene or the nucleotide sequence of claim 5 or claim 6 have about at least 94% sequence identity;
Ii. the 2nd RNA zone comprises these at least 19 the continuous nucleotide complementary nucleotide sequences with a RNA zone; With
Iii. the first and second RNA zones can base pairing, thereby forms double stranded rna molecule between these at least 19 continuous nucleotides in first and second zones;
F) transcribe the DNA that post-treatment becomes miRNA in vegetable cell, wherein this miRNA can discern the mRNA and the cutting of instructing this mRNA of the endogenous SusC encoding gene of plant.
15. the vegetable cell of claim 14 wherein can be expressed the effective described dna molecular that is connected of promotor with described plant and stably be incorporated into genome, in the cell nucleus gene group of preferred described vegetable cell.
16. the vegetable cell of claim 14 or claim 15, wherein said cell is from the fiber production plant.
17. the vegetable cell of claim 16, wherein said cell is from cotton.
18. comprise claim 14 to 17 each vegetable cell or the plant of forming by each vegetable cell of claim 14 to 17 basically.
19. the plant of claim 18, described plant are the fiber production plants.
20. the plant of claim 18, described plant is a vegetable lamb.
21. seed of the plant of each of claim 18 to 20 or progeny plants, it comprises the vegetable cell of claim 14.
22. fibre of plant from claim 20 or 21.
23. be used to change the method for fiber production fibre of plant feature, described method comprises sucrose synthase C isotype in the cell that changes described plant or cell walls or the apoplast liquid or has the functional level of the sucrose synthase of similar features.
24. the method for claim 23 wherein increases sucrose synthase C isotype or has the functional level of the sucrose synthase of similar features.
25. the method for claim 24 wherein increases the functional level of sucrose synthase C isotype by the expression cassette that claim 7 or claim 8 are provided to plant.
26. the method for claim 24, the functional level by provide expression cassette to increase the sucrose synthase similar to sucrose synthase C isotype to plant wherein, described expression cassette comprise and have the plant that the DNA zone of the sucrose synthase of hydrophobicity N end regions effectively is connected with coding and can express promotor.
27. the method for claim 23 wherein reduces sucrose synthase C isotype or has the functional level of the sucrose synthase of similar features.
28. the method for claim 27 wherein reduces the functional level of sucrose synthase C isotype by each the expression cassette that claim 9 to 13 is provided to plant.
29. the method for each of claim 23 to 28, a feature in the wherein following fiber characteristics is changed: intensity, length, fiber fineness, fiber maturity ratio, immature fibre content, fibre uniformity or mark are swelled Buddhist nun's value.
30. the albumen of each of claim 1 to 3 is used for changing the purposes of the fiber characteristics of fiber production plant.
31. the nucleic acid of claim 5 or claim 6 is used for changing the purposes of the fiber characteristics of fiber production plant.
32. the fiber that each the method by claim 23 to 29 obtains.
33. fabric (tissue) or yarn (yarn) that the fiber of Accessory Right requirement 22 or claim 32 is made.
34. be used for increasing the apoplast UDP-glucose of vegetable cell and the method for fructose concentration, this method comprises the expression cassette that claim 7 or 8 are provided to described vegetable cell.
CNA2007800286149A 2006-07-25 2007-07-20 Identification of a novel type of sucrose synthase and use thereof in fiber modification Pending CN101495626A (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
EP06015433 2006-07-25
EP06015433.3 2006-07-25
US60/834,066 2006-07-28

Publications (1)

Publication Number Publication Date
CN101495626A true CN101495626A (en) 2009-07-29

Family

ID=40925399

Family Applications (1)

Application Number Title Priority Date Filing Date
CNA2007800286149A Pending CN101495626A (en) 2006-07-25 2007-07-20 Identification of a novel type of sucrose synthase and use thereof in fiber modification

Country Status (3)

Country Link
CN (1) CN101495626A (en)
BR (1) BRPI0714552A2 (en)
ZA (1) ZA200900106B (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106834342A (en) * 2017-01-18 2017-06-13 华中农业大学 The method that Culm of Rice lodging tolerance is improved using sucrose synthase
CN110055232A (en) * 2019-01-10 2019-07-26 北京理工大学 Two Radix Glycyrrhizae sucrose synthases and its application in synthesis enoxolone glycosylated derivative
CN112280795A (en) * 2020-11-17 2021-01-29 昆明理工大学 Use of glycosyltransferase genes
WO2023226978A1 (en) * 2022-05-23 2023-11-30 苏州引航生物科技有限公司 Microbial-derived sucrose synthase and use thereof

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106834342A (en) * 2017-01-18 2017-06-13 华中农业大学 The method that Culm of Rice lodging tolerance is improved using sucrose synthase
CN110055232A (en) * 2019-01-10 2019-07-26 北京理工大学 Two Radix Glycyrrhizae sucrose synthases and its application in synthesis enoxolone glycosylated derivative
CN110055232B (en) * 2019-01-10 2020-08-25 北京理工大学 Two glycyrrhetinic acid sucrose synthases and application thereof in synthesis of glycyrrhetinic acid glycosylated derivatives
CN112280795A (en) * 2020-11-17 2021-01-29 昆明理工大学 Use of glycosyltransferase genes
WO2023226978A1 (en) * 2022-05-23 2023-11-30 苏州引航生物科技有限公司 Microbial-derived sucrose synthase and use thereof

Also Published As

Publication number Publication date
ZA200900106B (en) 2010-03-31
BRPI0714552A2 (en) 2013-05-07

Similar Documents

Publication Publication Date Title
EP1349446B1 (en) Modification of sucrose synthase gene expression in plant tissue and uses therefor
CN100575490C (en) Change the ways and means that produces fiber characteristics in the textile plant
US9121074B2 (en) Gossypium hirsutum plants with increased fiber strength comprising a fiber strength allele spanning the GLUC1.1A gene from Gossypium barbadense
Li et al. GbEXPATR, a species‐specific expansin, enhances cotton fibre elongation through cell wall restructuring
Hussey et al. SND2, a NAC transcription factor gene, regulates genes involved in secondary cell wall development in Arabidopsis fibres and increases fibre cell area in Eucalyptus
CN101578371B (en) Differential expression of subgenome specific alleles in cotton and uses thereof
Weschke et al. The role of invertases and hexose transporters in controlling sugar ratios in maternal and filial tissues of barley caryopses during early development
CN103403016A (en) Spatially modified gene expression in plants
CN101495626A (en) Identification of a novel type of sucrose synthase and use thereof in fiber modification
Li et al. Overexpression of UDP-glucose dehydrogenase from Larix gmelinii enhances growth and cold tolerance in transgenic Arabidopsis thaliana
US20100167040A1 (en) Identification of a novel type of sucrose synthase and use thereof in fiber modification
Wisniewski et al. Isolation of lipoxygenase cDNA clones from pea nodule mRNA
US7317136B1 (en) Methods for modifying plant cell walls and modified plants produced thereby
KR20200070357A (en) Plant resistance
CN111218454B (en) GhRFP1 gene and recombinant vector thereof
US7498492B2 (en) Modification of sucrose synthase gene expression in plant tissue and uses therefor
AU2002220363B2 (en) Modification of sucrose synthase gene expression in plant tissue and uses therefor
CN104650205A (en) Protein for controlling rice leaf morphology and root hair development, and coding gene and application thereof
ZA200303930B (en) Modification of sucrose synthase gene expression in plant tissue and uses therefor.
AU2002220363A1 (en) Modification of sucrose synthase gene expression in plant tissue and uses therefor
CN116103330A (en) Application of GhACY gene in improving drought resistance and yield of cotton
Holaday et al. Transgenic Cotton Over-Producing Spinach Sucrose Phosphate Synthase Showed Enhanced Leaf Sucrose Syntheis and Improved Fiber Quality Under Controlled Environmental Conditions

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C02 Deemed withdrawal of patent application after publication (patent law 2001)
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20090729