CN102094027B - Rice blast resistance gene Pi7 and application thereof - Google Patents
Rice blast resistance gene Pi7 and application thereof Download PDFInfo
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- CN102094027B CN102094027B CN2010105786332A CN201010578633A CN102094027B CN 102094027 B CN102094027 B CN 102094027B CN 2010105786332 A CN2010105786332 A CN 2010105786332A CN 201010578633 A CN201010578633 A CN 201010578633A CN 102094027 B CN102094027 B CN 102094027B
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Abstract
The invention discloses a rice blast resistance gene Pi7 and application thereof, and provides a nucleotide sequence of the rice blast resistance gene Pi7 and an amino acid polypeptide sequence coded thereby. The nucleotide sequence comprises two genes which belong to members of a coiled-coil nucleotide binding site leucine-rich repeat (CC-NBS-LRR) resistance gene family and are constitutive expression genes. The invention also provides application of the gene to the aspect of transforming rice or breeding disease-resistant varieties of other plants and application of a molecular marker produced according to the gene sequence to breeding.
Description
Technical field
The invention belongs to gene engineering technology field, be specifically related to a kind of resistance gene of rice blast
Pi7Clone and application thereof.
Background technology
Plant usually receives the infringement of the several diseases original in the process of growth, plant then takes multiple defence policies to protect self, avoids receiving it to attack.A most important defense mechanism is exactly the defence answering system that can discern the existence of obligate pathogenic microorganism and start self in the plant.Plant is mediated by disease-resistant gene the identification of pathogenic bacteria.Therefore, enantiopathy gene product structure is analyzed and is studied, and is the basis of understanding plant disease-resistant mechanism, also has important directive significance for the prevention and the control of its disease.
Up to now, oneself is through having separated more than 80 disease-resistant gene from plant.To the structure of these disease-resistant genes and discovering of product, though host plant is different, pairing pathogen also has differences such as fungi, bacterium, virus and nematode; But the structure of disease-resistant gene and product have many common constitutional featuress, as C-end exist rich leucine Tumor-necrosis factor glycoproteins (leucine-rich repeat, LRR); N-end exist nucleotide binding site (nucleotide binding site, NBS), leucine zipper (leucine zipper; LZ), and the coiled coil structural domain (coiled-coil, CC); Membrane spaning domain (transmembrane domain, TM), protein kinase (protein kinase; PK), and fruit bat Toll albumen and Mammals interleukin-1 acceptor (Toll and interleukin – 1 receptor, TIR) etc.According to they coded proteic constitutional featuress, can disease-resistant gene be divided into 7 types of (Hammond-Kosack & Jones, 1997; Dangl & Jones 2001; Iyer & McCouch 2004).
The first kind, toxin reduction enzyme disease-resistant gene.Like the corn disease-resistant gene
Hm1, it is the 1st plant disease resistance genes that quilt is cloned, it is controlled fungi
Cochliobolus carbonumThe resistance of microspecies 1.
Hm1Coding separate toxenzyme can passivation the HC toxin that produced of pathogenic fungi, and the HC toxin is a fungi
C.
CarbonumThe virulence factor that microspecies 1 produce, it determines some genotype (Johal etc., 1992) that this germ can only infect corn.Second type, NBS-LRR class disease-resistant gene.The nearly N end of their encoded protein is NBS, and nearly C end then is made up of LRR.As
RPS2(Bent et al., 1994),
RPM1(Grant et al., 1995),
I2(Simon et al., 1998);
RPP5(Parker et al., 1997),
N(Dodd et al., 2001),
L 6(Lawrence et al., 1995),
Mla1(Zhou et al., 2001),
Mla6(Halterman et al., 2001); The disease-resistant gene of paddy rice as
Xa1(Yoshimura et al., 1998),
Pib(Wang et al., 1999),
Pita(Bryan et al., 2000) etc.The 3rd type, PK class disease-resistant gene.Like tomato
PtoGene, its product are a kind of intracellular serine-threonine protein kinase enzymes that is positioned at, and do not have LRR structural domain (Martin et al., 1993).The 4th type, LRR-TM class disease-resistant gene.The gene of the different physiological strains of the anti-leaf mold of tomato
Cf-2(Dixon et al., 1996),
Cf-4(Thomas et al., 1997),
Cf-5(Dixon et al., 1998),
Cf-9(Jones et al., 1994), and the gene of the anti-SCN of beet
Hs1 Pro-1 (Cai et al., 1997) etc.The 5th type, LRR-TM-PK class disease-resistant gene is with the rice bacterial blight resistance gene
Xa21(Song et al., 1995) are representative.The 6th type, with Arabidopis thaliana
RPW8Be representative, its encoded protein only contains complete CC and NBS structural domain (Xiao et al., 2001).The 7th type, with paddy rice
Xa5Gene is representative, and its encoded protein is a transcription factor (TFIIA γ) (Iyer & McCouch, 2004).
The disease-resistant proteic disease-resistant gene of coding NBS-LRR class is one type of maximum in plant disease resistance genes disease-resistant gene; Constructional feature according to the disease-resistant protein N terminal of NBS-LRR class; Can this genoid be divided into two big types of TIR-NBS-LRR and CC-NBS-LRR (Meyers et al., 2002 again; Pan et al., 2000; Cannon et al., 2002; Richly et al., 2002).TIR-NBS-LRR class disease-resistant gene is mainly found in dicotyledons, also in the monocotyledons genome, does not find (Bai et al., 2002 so far; Meyers et al., 2002).The disease-resistant gene that in monocotyledons, identifies the at present disease-resistant albumen of CC-NBS-LRR class of mainly encoding; Also there is a large amount of CC-NBS-LRR class disease-resistant genes in the dicotyledons; Comparatively speaking; CC-NBS-LRR class disease-resistant gene in the monocotyledons is than more rich and varied (Cannon et al., 2002) in the dicotyledons.
Research shows; NBS, CC, TIR structural domain possibly participated in signal conduction (Hammond-Kosack & Jones 1997); Although this type R albumen does not have the intrinsic kinase activity; But NBS can activate kinases or G albumen, and NBS has the ATP of combination or GTP and hydrolytic enzyme activities (Traut et al., 1994).The signal conduction (Jones et al., 1994) in the disease-resistant defense response of TIR structure possibility involved in plant downstream.Nearest evidence shows, flax
LFamily's variety selects to also occur in the TIR zone, and this zone forms specificity (Luck et al., 2000) with corresponding LRR zone coevolution.The function of LRR structural domain be mainly concerned with protein-protein and with interaction (Jones & Jones, 1996 of part; Kajava et al., 1998), be product or the indirect interactional position of product (Bent, 1996 that the disease-resistant gene product is direct and the pathogenic bacteria nontoxic gene is encoded by inference; Baker et al., 1997).Jia et al. (2000) proves through yeast two-hybrid
AVR-PitaEncoded protein is processed as one 176 amino acid whose activated protein AVR-Pita
L76The specific exciton of microspecies is delivered to plant cytoplasm and combines with the LRD zone of Pita acceptor specifically, thus the defense response of Pita mediation in the activating cells.Av-rPita after the Ala among the Pita becomes Ser
176Can not combine with LRD, thereby show susceptible.This result has proved that directly the LRR structural domain possibly be exactly the zone of pathogenic bacteria identification; Mutual " gene pairs gene " relation of making to have verified from molecular level for the first time paddy rice and Pyricularia oryzae of Pita and AvrPita.
Paddy rice is one of most important food crop in the world, and it is staple food with rice that population over half is arranged approximately.By pathogenic fungi
Magnapothe oryzae(no condition:
Pyricularia oryzae) rice blast that causes is that Rice Production is endangered one of severe diseases, annually all causes serious grain loss.From the viewpoint of Sustainable development of environment protection with agricultural, breed and the utilization of disease-resistant variety is the safe and effective procedure of control rice blast.But; Because the variety and the volatility of rice blast fungus population; People lack effective utilization of antagonism gene in addition; And antagonism mechanism lacks fully and understands, so that susceptibleization of disease-resistant variety problem not only is not resolved, and becomes thremmatology man stubborn problem the most because of the shortage of effective anti-source gene and short-livedization of disease-resistant variety on the contrary.Therefore, excavate, identify and clone disease-resistant gene and reasonably be applied to breeding for disease resistance in the works, become the preferential major issue that solves in the agri-scientific research.
Along with molecular biological fast development, so far, have at least more than 80 rice blast master to imitate resistant gene and be in the news, wherein located for 60 by molecule.At present; On the 3rd karyomit(e) of paddy rice, also not identified the rice blast master imitates the resistant gene; All identified the main resistant gene site of imitating on remaining 11 karyomit(e); And contain a plurality of rice blast resistances site on the karyomit(e) that has, and the resistant gene of some gene locus is that cluster exists.At present, aspect clone's research of rice blast resistance gene, existing 13 resistant genes of formal report,
Pib(Wang et al., 1999),
Pita(Bryan et al., 2000),
Pi9(Qu et al., 2006),
Pid2(Chenet al., 2006),
Piz-tWith
Pi2(Zhou et al
., 2006b),
Pi36(Liu et al., 2007a),
Pi37(Lin et al., 2007),
Pik-m(Ashikawa et al., 2008),
Pit(Hayashi et al
.,2009),
Pi5(Lee et al
.,2009),
Pid3(Shang et al
.,2009) and
Pi21(Fukuoka et al., 2009).
Clone's disease-resistant gene is the prerequisite to the further investigation of paddy rice resistance mechanism, discloses the molecule mechanism of rice anti-rice blast and can control and reduce the harm of Pyricularia oryzae to paddy rice better.Simultaneously, clone's disease-resistant gene is modified and transformed, can control and increase the disease resistance of plant artificially, widen the anti-spectrum of plant.These aspects are that employing conventional plant breeding and improving technology institute are inaccessiable.
Summary of the invention
The purpose of this invention is to provide a kind of resistance gene of rice blast and the dna fragmentation that comprises the promotor of regulating and control this gene.
Another object of the present invention provides the coded protein of above-mentioned rice blast resistance gene.
Another object of the present invention provides the above-mentioned carrier that contains above-mentioned resistant gene.
Another object of the present invention provides the host cell of above-mentioned carrier.
Another object of the present invention provides the application of said gene in the preparation transgenic plant.
Further purpose of the present invention provides molecule marker that above-mentioned resistant gene produces and rice blast is had the application in the paddy rice of disease resistance in seed selection.
The present invention separates from rice varieties IRBL7-M and obtains
Pi7Gene comprises the gene of 2 coding NBS-LRR proteinoids
Pi7-1With
Pi7-2, its nucleotide sequence shown in SEQ ID NO.1 or SEQ ID NO.2, their aminoacid sequences shown in SEQ ID NO.3 and the SEQ ID NO.4 of encoding respectively, structure like accompanying drawing 9 with shown in the accompanying drawing 10.Their albumen all comprises 2 main structural domain: NBS and LRR zone, wherein
Pi7-1The NBS structural domain contains conservative kinase 1a:GLPGGGKTTVAR, kinase2:NKKYLIVIDDIW, kinase3a:DLGGRIIMTTRLNSI, GLPL:EDSPCYDIVNMCYGMPLALIW;
Pi7-2The NBS structural domain contains conservative kinase1a:VLSIVGFGGVGKTTIA; Kinase2:LEQLLAEKSYILLIDDIW, kinase3a:GGRIIVTTRFQAV, GLPL:EQVPEEIWKICGGLPLAIV; RNBS-D:CLLYLSIFPKGWK, MHDV:KTFQVHDMVLEYI.And the proteic C-end of Pi7-1 is 16 incomplete LRR repetitions, and its leucine content is 14.0% (Fig. 9); The proteic C-end of Pi7-2 is that 13 incomplete LRR repeat, and its leucine content is 17.0% (Figure 10).
Be to be understood that; Under the prerequisite that does not influence protein-active (not in proteic active site), those skilled in the art can carry out various replacements, interpolation and/or lack the aminoacid sequence that one or several amino acid obtains to have same function the aminoacid sequence shown in SEQ ID NO.3 or the SEQ ID NO.4.In addition, consider the degeneracy of codon, for example can be in its coding region, under the condition that does not change aminoacid sequence, or at its non-coding region under the condition that does not influence protein expression, above-mentioned proteic gene order is made amendment to encoding.Therefore, the present invention also comprises replacement, the interpolation that the above-mentioned proteic gene order of encoding is carried out and/or lacks one or more Nucleotide, has the nucleotide sequence that has identical function with above-mentioned encoding sox.The present invention also comprises just sequence or the antisense sequences based on said gene, comprises the host cell that contains said nucleotide sequence or its segmental cloning vector or expression vector, contain said carrier, contains said nucleotide sequence or its segmental plant transformed cell and transgenic plant.
The present invention comprise equally with
Pi7The primary structure part of resistant gene connects the upward formed mosaic gene of proper regulation sequence effectively, and in genome, comprises the plant of this gene and the seed of this kind of plant.This gene can be natural or chimeric.For example, will comprise the fragment of this gene and the promotor of a constitutive expression and be connected, this promotor can be expressed with cytocerastic any period under any condition.The promotor of this constitutive expression comprises the 35S promoter of cauliflower mosaic virus etc.On the other hand, promotor that also can the promotor of this gene and a tissue specific expression or developmental stage is specific expressed or accurately the promotor of environmental induction be connected, these promotors are referred to as inducible promoter.Like this, the change of environment, the difference of developmental stage can change this expression of gene.Likewise, also can this expression of gene be limited in some tissues, make by this gene induced disease resistance response and obtain artificial control.Wherein envrionment conditions comprises attack, anaerobic condition and the light etc. of pathogenic bacteria, and tissue and developmental stage comprise leaf, fruit, seed and flower etc.
The present invention also further clones the promotor that obtains said gene, and the dna fragmentation that comprises the promotor of regulating and control this gene is respectively shown in SEQ ID NO.5 and SEQ ID NO.6.
According to provided by the invention
Pi7Gene order information (SEQ ID No.1 and SEQ ID No.2), those skilled in the art can through following method easily obtain with
Pi7The gene that is equal to: (1) obtains through database retrieval; (2) with
Pi7Gene fragment is that genomic library or the cDNA library of probe Screening of Rice or other plant obtains; (3) basis
Pi7Gene order information design oligonucleotides primer obtains from genome, mRNA and the cDNA of paddy rice or other plant with the method for pcr amplification; (4) exist
Pi7Obtain with the gene engineering method transformation on the basis of gene order; (5) method with chemosynthesis obtains this gene.
Rice blast resistance gene provided by the invention
Pi7Have important use and be worth, its give plant to Pyricularia oryzae (
Magnaporthe oryzae) caused disease produces specific disease resistance response.It is applicable to that these plants comprise monocotyledons and dicotyledons to all plants responsive to this pathogenic bacteria.One of application is with described
Pi7Gene order is connected to any plant conversion carrier, will with any method for transformation
Pi7Disease-resistant gene imports paddy rice or other plant cell, can obtain the transgenic disease-resistant variety of expressing said gene, thereby is applied to produce.Of the present invention gene constructed in plant conversion carrier; Can do suitable modification to said gene or its regulating and controlling sequence; Also can before its transcription initiation codon, replace the original promotor of said gene, thereby widen and the resistance of enhancement of plant pathogenic bacteria with other promotor.
The Another application of resistant gene provided by the invention is according to the specific molecule marker of said gene order information generating, includes but not limited to SNP (mononucleotide polymorphic), SSR (simple sequence repeats polymorphic), RFLP (restriction enzyme length is polymorphic), CAPS (the cutting amplified fragments is polymorphic).Can identify the resistant gene type of paddy rice or other plant with this mark, be used for the molecular marker assisted selection breeding, thereby improve the efficiency of selection of breeding.
The present invention has tangible advantage and effect.Change clone's disease-resistant gene over to susceptible plant, help to produce new disease-resistant plants.Particularly can be with transformation technology a plurality of disease-resistant genes that in plant, add up, and can not produce the chain problem of bad gene in the genome of following appearance in the traditional breeding technology, and can shorten breeding time.The clone of disease-resistant gene overcomes the prerequisite that can not between plant species, shift the problem of disease-resistant gene in the traditional breeding method.
The present invention can further provide or use the disease-resistant transfer-gen plant and corresponding seed that obtains based on above-mentioned dna fragmentation, and with gene of the present invention or based on the recombinant chou plants transformed of this gene or the seed that obtains by this type plant.Can gene of the present invention be changed in other the plant with the mode of sexual hybridization.
Description of drawings
Fig. 1 is a resistance gene of rice blast
Pi7Clone's policy map.Before there had been report to claim
Pi1(C101LAC),
Pi7(RIL29) and
PikTo go out identical response type (Tsunematsu et al. 2000, Breeding Science 50:229-234) from Filipine a plurality of Pyricularia oryzae strains expressed;
Pi1With
Pi7In fact be
PikAllelotrope or gene identical (Campbell et al. 2004, Phytopathology 94:302-307) with it.In conjunction with this research department pair with above-mentioned
PikAllelotrope has " gene pairs gene, gene-for-gene " interactional avirulence gene of rice blast location, clone's result, thereby infers
Pi7Possibly be exactly
PikNew allelotrope.Therefore, the present invention is to have announced
Pi7Localized information (Campbell et al. 2004, Phytopathology 94:302-307) is the basis, makes up the electronics physical map in this goal gene zone; Adopt with
PikAllelotrope it
Pik-m(Ashikawa et al. 2008, Genetics 180:2267 – 2276) and
Pik(Zhai et al. 2010, New Phytologist DOI:10.1111/j.1469-8317.2010.03462.x) similar clone's strategy is cloned this resistant gene.
Fig. 2 does
PikBunch allelotrope is to the comparison of the anti-spectrum of 8 rice blast fungus populations.Ordinate zou is a fastness frequency among the figure, and X-coordinate is 8 rice blast fungus populations (GD, Guangdong; FJ, Fujian; HN, the Hunan; GZ, Guizhou; SC, Sichuan; LN, Liaoning; JL, Jilin; HLJ, Heilungkiang).5
PikBunch allelotrope (IRBLks-S,
Pik-s; IRBLk-Ka,
Pik; Tsuyuake,
Pik-m; IRBLkp-K60,
Pik-p; IRBL7-M,
Pi7).This shows,
Pi7The higher resistance of rice blast fungus population performance to Guangdong, Hunan, Guizhou, Sichuan and Liaoning.
Fig. 3 is a goal gene
Pi7The design of graphics of zone electronics physical map.Fig. 3 A: above-mentioned Campbell etc. (2004) make up
Pi7The genetic map that the zone is rough; Fig. 3 B: according to
Pik-m(Ashikawa et al. 2008, Genetics 180:2267 – 2276) and
PikThe locating information of (Zhai et al. 2010, New Phytologist DOI:10.1111/j.1469-8317.2010.03462.x) makes up
Pi7With
PikBunch allelic integration physical map; Fig. 3 C: 4 BAC clones that select gene region to exist; Fig. 3 D:
PikGene candidate region (Zhai et al. 2010, New Phytologist 180:2267 – 2276); Fig. 3 E:
Pi76 disease-resistant candidate genes that gene locus exists on the fine type gene group of Japan with NBS-LRR conserved structure; Fig. 3 F:
Pi72 disease-resistant candidate genes that gene locus exists on IRBL7-M/Tsuyuake type gene group with NBS-LRR conserved structure.This shows,
Pi7There is two types genome on the gene locus, has very big disappearance/insertion each other;
Pi7Gene have only 2 candidate genes (
Pi7-1With
Pi7-2).
Fig. 4 is a resistance gene of rice blast
Pi7(by
Pi7-1With
Pi7-2Composition) genetic complementation T
0The resistance of plant is identified pictorial diagram.Solid arrow refers to disease-resistant plant among the figure, and disease plant that dotted arrow refers to is explained resistant gene
Pi7By
Pi7-1With
Pi7-2Forming could the function of appeal.
Fig. 5 is a resistance gene of rice blast
Pi7The part T of genetic complementation
0The PCR of the selective marker HPT gene of transformant detects figure.Among the figure from left to right, swimming lane 1,15: molecular weight marker DL2000; Swimming lane 2: empty carrier (Vector), positive control; Swimming lane 3: transformation receptor kind Q1063, negative control; Swimming lane 4: donor kind IRBL7-M, negative control; Swimming lane 5-14:T
0Transformant.The result shows; All disease-resistant transformant have all detected the pcr amplification product of selected marker; And susceptible transformant does not detect the pcr amplification product of selected marker, explains that disease-resistant transformant is to have recovered resistance owing to transformation construct has been incorporated into to have obtained to express in the genome of acceptor kind Q1063 of high sense.R: high anti-; MR: in anti-; MS: middle sense; S: susceptible.
Fig. 6 is a resistance gene of rice blast
Pi7Genetic complementation offspring's genotype and phenotypic coseparation analysis figure.Among the figure from left to right, swimming lane 1,11: molecular weight marker DL2000; Swimming lane 2: empty carrier (Vector), positive control; Swimming lane 3: transformation receptor kind Q1063, negative control; Swimming lane 4-10:T
1Transform the separation offspring of system.The result shows, at T
1Transform among the separation offspring of system the T that all are disease-resistant
1Transformant has all detected the pcr amplification product of selected marker, and susceptible T
1Transformant does not detect the pcr amplification product of selected marker, and this T is described
1Conversion is that the individual genotype of offspring is divided into phenotype and leaves, that is transgenic is separated genetic stability expression among the offspring at it.R: high anti-; MR: in anti-; S: susceptible.
Fig. 7 is a resistance gene of rice blast
Pi7RNA interfere (RNAi) F
1The resistance of plant is identified pictorial diagram.Among the figure HF4 be donor kind IRBL7-M with
Pik-h(be
Pi7Height homologous allelotrope) RNAi T
1The F of transformant kh4i-11 genetic cross
1Plant, its postvaccinal performance disease is susceptible (S), explanation
Pi7The function quilt
Pik-hRNAiT
1The construct that transformant contained and interfered (destruction); HF6 be donor kind IRBL7-M with
Pik-hRNAiT
1The F of transformant kh4i-26 genetic cross
1Plant, its postvaccinal performance disease is susceptible, explanation
Pi7The function quilt
Pik-hRNAiT
1The construct that transformant contained and interfered (destruction); HF10 is for containing among the figure
PiaKind Aichi Asahi with
Pik-h(
PiaBe with
Pik-h/
Pi7The non-allelic genes that the position is different) RNAi T
1The F of transformant kh4i-9 genetic cross
1Plant, its postvaccinal phenotype are disease-resistant (R), explanation
PiaThe resistance function do not have quilt
Pik-hRNAiT
1The construct that transformant contained and interfered (destruction) therefore, can be used as the contrast that non-specific RNA interferes; IRBL7-M is
Pi7The donor kind, its postvaccinal phenotype is disease-resistant (R), explain the inoculation used rice blast fungi isolates CHL381 be suitable.
Fig. 8 is a resistance gene of rice blast
Pi7RNA interfere (RNAi) F
1The Molecular Identification pictorial diagram of plant.Among the figure from left to right, swimming lane M (both sides): molecular weight marker DL2000; Swimming lane 1: above-mentioned IRBL7-M, do not detect the special molecule marker of RNAi construct (
Hpt), promptly do not contain construct; Swimming lane 2:RNAi T
1Transformant kh4i-11, detected the special molecule marker of RNAi construct (
Hpt), promptly contain construct; Swimming lane 3: the F of above-mentioned HF4
1Plant contains construct; Swimming lane 4:RNAi T
1Transformant kh4i-26 contains construct; Swimming lane 5,6: the F of above-mentioned HF6
1Plant contains construct; Swimming lane 7: above-mentioned Aichi Asahi, do not contain construct; Swimming lane 8:RNAi T
1Transformant kh4i-9 contains construct; Swimming lane 9,10: the F of above-mentioned HF10
1Plant contains construct.Above presentation of results, the F of HF4 and HF6
1Plant make because receive allele specific RNA to interfere its resistant gene that contains (
Pi7) function do not express; As contrast, the F of HF10
1Plant make because the allele specific RNA of the right and wrong that receive interferes its resistant gene that contains (
Pia) the function normal expression.
Fig. 9 is a resistance gene of rice blast
Pi7Constitutive gene
Pi7-1Gene structure and sequence chart.Fig. 9 A: light box indicating 5 ' and 3 '-UTR among the figure; Dark box indicating exon; Lines are represented intron; Promotor of this gene coding region (ATG) and terminator (TAG) and each regional size (bp) also are shown in wherein.Fig. 9 B: the amino acid of CC motif represented to form in the boldface type of CC structural domain band underscore among the figure, and aminoacid sequence conservative among the NBS represented in the boldface type of NBS zone band underscore.The amino-acid residue that independently list the lower section is the terminal LRR of C-zone; The conservative motif of xLDL in LRR zone represented to form in the boldface type of band underscore; Back, LRR zone also has one section CNL (C holds non-LRR zone) aminoacid sequence.Two aminoacid sequences that indicate that roll off the production line do
Pi7Special amino acid replacement [substitution, with
PikBunch other allelotrope (
Pik,
Pik-s,
Pik-p,
Pik-m) compare].
Figure 10 is a resistance gene of rice blast
Pi7Another constitutive gene
Pi7-2Gene structure and sequence chart.Figure 10 A: light box indicating 5 ' and 3 '-UTR among the figure; Dark box indicating exon; Lines are represented intron; Promotor of this gene coding region (ATG) and terminator (TAG) and each regional size (bp) also are shown in wherein.Figure 10 B: the amino acid of CC structure represented to form in the boldface type of CC structural domain band underscore among the figure, and aminoacid sequence conservative among the NBS represented in the boldface type of NBS zone band underscore.The amino-acid residue that independently list the lower section is the terminal LRR of C-zone; The conservative motif of xLDL in LRR zone represented to form in the boldface type of band underscore.Two aminoacid sequences that indicate that roll off the production line do
Pi7Special amino acid replacement [substitution, with
PikBunch other allelotrope (
Pik,
Pik-s,
Pik-p,
Pik-m) compare].
Figure 11 is a resistance gene of rice blast
Pi7The quantitative RT-PCR of expression characterization detects figure.Left side figure is a constitutive gene
Pi7-1Different inoculation time points (0 hr, 12 hr, 24 hr, 72 hr, relative expression's level 120hr) this shows, this expression of gene type does, progressively raises from 12 hr-72 hr, then progressively downward modulation.Right figure is another constitutive gene
Pi7-2Relative expression's level at different inoculation time points this shows, this expression of gene type is that 12 hr-24 hr progressively raises, then progressively downward modulation.It can also be seen that thus this gene is at inoculation contrast (H
2O) and between the inoculation pathogenic bacteria show visibly different inducible expression.2 genes all show as constitutive expression because the inoculation before with can both detect 2 expression of gene afterwards.Ordinate zou is relative expression's level among the figure, and X-coordinate express time, unit are hour (h), and the twill block diagram is contrast (before the inoculation), and black surround figure is inoculation contrast (H
2O), blank block diagram is the pathogenic bacteria inoculation.
Figure 12 is for utilizing resistance gene of rice blast
Pi7The genotype identification figure of specific molecular marker.This shows, the combination of 2 specific molecular markers use can with
Pi7Distinguish with other several allelotrope.Figure 12 A is mark T1-795G, this mark can with the genotype of resistant variety IRBL7-M (
Pi7/Pi7) and resistant variety IRBLkp-K60 (
Pik-p/
Pik-p) and susceptible acceptor kind dragon round-grained rice 24Q1063 (
Pi7/pi7) distinguish; Figure 12 B is mark A2-1879G, this mark can with the genotype of resistant variety IRBL7-M (
Pi7/Pi7) and resistant variety IRBLk-Ka (
Pik/Pik), IRBLkm-Ts (
Pik-m/Pik-m), IRBLks-S (
Pik-s/Pik-s), susceptible acceptor kind Q1063 (
Pi7/pi7), susceptible acceptor kind dragon round-grained rice 24 (
Pi7/pi7) distinguish.Among the figure, M: molecular weight marker DL2000.
Embodiment
Following examples further specify content of the present invention, but should not be construed as limitation of the present invention.Under the situation that does not deviate from the present invention's spirit and essence, modification or replacement to the inventive method, step or condition are done all belong to scope of the present invention.If do not specialize the conventional means that used technique means is well known to those skilled in the art among the embodiment.In the embodiments of the invention part, we have set forth
Pi7Resistance characteristic, separating clone, functional character and the Molecular Identification of gene, isolating
Pi7Gene can be connected with appropriate carriers, changes in the plant materials, makes this plant materials have certain resistance.
In order to compare with clear and definite
Pi7With
Pik4 allelotrope in site (
Pik,
Pik-p,
Pik-s,
Pik-m) anti-spectrum, utilize from Guangdong (60 bacterial strains) Fujian (40); Hunan (41), Guizhou (60), Sichuan (66); Liaoning (108), 498 bacterial strains of total that Jilin (60) and Heilungkiang (63) collect to above-mentioned 5 allelic kinds of holding (be respectively IRBL7-M, IRBLk-Ka; IRBLkp-K60, IRBLks-S and Tsuyuake) carried out anti-spectrum comparative analysis.The result shows,
Pi7To the higher resistance of rice blast fungus population performance in Guangdong, Hunan, Guizhou, Sichuan and Liaoning, explain that thus this gene can use (Fig. 2) in above-mentioned area.Rice varieties RBL7-M; IRBLk-Ka, IRBLkp-K60 and IRBLks-S document (Kobayashi et al. Development of new sets of international standard differential varieties for blast resistance in rice (
Oryza sativaL.). JARQ, 2007, open in 41:31-37).Rice varieties Tsuyuake is at document (Wang et al. Characterization of rice blast resistance genes in the
PikCluster and fine mapping of the
Pik-pLocus. Phytopathology, 2009, open in 99:900-905).
Previous Position Research shows
Pi7The assignment of genes gene mapping in
PikThe genome area that cluster gene is identical (Campbell et al. Development of co-dominant amplified polymorphic sequence markers in rice that flank the
Magnaporthe griseaResistance gene
Pi7(t) in recombinanant inbred line 29. Phytopathology, 2004,94:302-307; Fig. 3 A); The anti-spectrum analysis of this research department shows
Pi7Gene with
PikBunches of 3 allelotrope (except resistance more weak
Pik-sOutside) most of test strain is shown identical resistance reaction (Fig. 2); The previous research in this research department shows, with
Pi7Corresponding nontoxic gene
AvrPi7Also be with
PikThe nontoxic gene that bunch resistant gene is corresponding
AvrPik/km/kpBe homotopic.The result of comprehensive above-mentioned three aspects according to " gene pairs gene " theory of Plant diseases system, infers
Pi7Should also be
Pik/km/kpAllelotrope.Therefore, the present invention has adopted the clone with this research department
PikStrategy that allelotrope is similar and method have been accomplished
Pi7The structure (Fig. 1) of target area electronics physical map.That is, be the basis with paddy rice with reference to the sequence information of kind Japan fine (Nipponbare), through the information biology means, with
Pi7Chain mark (Campbell et al. 2004; Fig. 3 A) and with
PikGene is isolating mark (Zhai et al. 2010, New Phytologist DOI:10.1111/j.1469-8317.2010.03462.x altogether; Fig. 3 B) carry out physical location and integrate, and made up the contig figure (Fig. 3 C) in goal gene zone thus, and electronics physical map (Fig. 3 D).
In order to confirm
Pi7Candidate gene; The applicant utilizes the fine reference sequences of Japan; Through 3 kinds of predictive genes software RiceGAAS (http://ricegaas.dna.affrc.go.jp), Gramene (http: // 143.48.220.116/resources/) FGENESH (http://www.softberry.com) with Softberry has carried out predictive genes and note analysis to the goal gene zone, tentatively confirmed
Pi7Candidate resistance gene be 6 nucleotide binding sites and rich leucine repeat (nucleotide binding site-leucine-rich repeat, candidate gene NBS-LRR) (
Pi71-NP,
Pi72-NP,
Pi73-NP,
Pi74-NP,
Pi75-NPAnd
Pi76-NP).
To 6 candidate genes carry out based on the gene specific mark existence/disappearance (presence/absence P/A) analyzes, and the result shows, with
Pik Bunch 3 allelotrope are the same, 4 candidate genes that exist at the fine reference sequences of Japan (
Pi71-NP,
Pi72-NP,
Pi73-NP,
Pi74-NP)
Pi7The institute hold among the kind IRBL7-M be non-existent (Fig. 3 E, 3F).Therefore,
Pi7Candidate resistance gene have only
Pi7-1With
Pi7-2, next step will be through following its function of functional complementation experimental verification.
Previous research shows,
PikBunch 2 allelotrope
Pik-m(Ashikawa et al. 2008, Genetics 180:2267 – 2276) and
Pik(Zhai et al. 2010, New Phytologist DOI:10.1111/j.1469-8317.2010.03462.x) is all by 2 closely linked genomic constitutions.Therefore, utilize the high-fidelity enzyme
PhusionIn conjunction with long segment PCR (long-range PCR; LR-PCR) technology; Total DNA with IRBL7-M is a template, according to or with reference to Liu et al. (2007, Genetics; 176:2541-2549) with Zhai et al. (2010; New Phytologist DOI:10.1111/j.1469-8317.2010.03462.x) experimental technique and the program described are made as a whole gene with above-mentioned 2 candidate genes and have been carried out amplification, clone, have carried out then based on the acquired experiment of the function of genetic complementation (Fig. 1).The result shows, in the genetic complementation offspring, has obtained the transformant (table 1 that has complementary functions; Fig. 4).
Amplification, clone
Pi7-12Primer be:
Forward primer: TTTT
GGCGCGCCGGTGCGGGAAGAAGACGGGATAAAGA
Reverse primer: TTTT
GGCGCGCCGCCGACGATAGCAACCGACAGGAGTG
Utilize hygromycin gene in the conversion carrier (
Hpt) mark carried out the PCR detection to transformant.Detected result proof target gene fragment has imported in the disease-resistant transformant, and does not import (Fig. 5) in the susceptible transformant.
Table 1.
Pi7The genetic complementation experiment analysis results of 2 constitutive genes
aT from each construct
0The plant inoculation is right
Pi7Avirulent bacterial strain CHL346.R, disease-resistant; MR, in anti-; MS, middle sense; S, susceptible.
bThe success ratio that has complementary functions (%) is R+MR/R+MR+MS+S*100%
The experimental result of above-mentioned genetic complementation shows, candidate gene
Pi7-12Be exactly
Pi7Functional gene.Rice varieties Q1063 and bioabsorbable carrier material pCAMBIA1300 are open in document (Lin et al.2007, Genetics 177:1871-1880); Rice varieties Longjing 24 (imperial round-grained rice 24) is open in document (Zhai et al. 2010, New Phytologist DOI:10.1111/j.1469-8317.2010.03462.x).
In order further to confirm the genetic stability of transgenic constructs, select the transformation generation T of part at transformation generation
1Strain is to have carried out the coseparation analysis (Fig. 6) of resistance phenotype/resistant gene type.The result shows that resistance phenotype and resistant gene type are divided into fully and leave.That is transgenic constructs can genetic stability in transformation generation, expression.
For further affirmation
Pi7Function, and
Pi7Whether be exactly
PikBrand new allelotrope, the applicant carried out based on
PikAllelotrope (
Pik-h) T of RNAi
1The afunction property experiment (Fig. 1) of transformant filial generation.The result shows,
Pik2 plant (kh4i-11 and kh4i-26) that allelotrope RNAi isozygotys stable and the hybridization F of donor kind IRBL7-M
1Plant all shows
Pi7The forfeiture of resistance function.That is,
Pi7The resistance function because quilt
PikThe T of allelotrope RNAi
1The construct that transformant contained and interfered (destruction) (HF4 and HF6 among Fig. 7).Contrast as non-specific RNA interferes contains
PiaKind Aichi Asahi with
Pik-h(
PiaBe with
Pik-h/
Pi7The non-allelic genes that the position is different) RNAi T
1The F of transformant kh4i-9 genetic cross
1Plant, its postvaccinal phenotype still are disease-resistant (HF10 among Fig. 7), explanation
PiaThe resistance function do not have quilt
Pik-hRNAiT
1The construct that transformant contained and interfere (destruction).IRBL7-M is
Pi7The donor kind, its postvaccinal phenotype is disease-resistant, explain the inoculation used rice blast fungi isolates CHL381 be suitable.
For the forfeiture of confirming filial generation resistance function whether with RNAiT
1The construct that transformant contained is relevant, and the parent and the offspring thereof of above-mentioned 3 cross combinations carried out the Molecular Identification based on the construct specific molecular marker.The result shows; Offspring (HF4 and HF6) the resistance phenotype relevant with its molecular marker gene type ( swimming lane 3,5,6 of Fig. 8) that specific RNA is interfered, but not offspring (HF10) the resistance phenotype uncorrelated with its molecular marker gene type ( swimming lane 9,10 of Fig. 8) that specific RNA is interfered.
The employing method of moving one's steps is right
Pi7Dna sequence dna measure.Utilize the RACE technology, obtained
Pi7Full length cDNA sequence and it is checked order.
Pi7Gene DNA length is that 16.9 kb (comprise
Pi7-1With
Pi7-2), wherein,
Pi7-1Full length cDNA sequence be 3723 bp, contain the opening code-reading frame of one 3429 bp, 5 ' and 3 ' non-translational region is respectively 104 bp and 190 bp.Through icp gene group DNA and cDNA sequence, find that the opening code-reading frame of this gene contains 2 introns and 3 exons (Fig. 9 A).Another constitutive gene
Pi7-2Full-length cDNA be 3690 bp, contain the opening code-reading frame of one 3066 bp, 5 ' and 3 ' non-translational region is respectively 380bp and 244bp.Through icp gene group DNA and cDNA sequence, find that the opening code-reading frame of this gene contains 1 intron and 2 exons, and contain an intron (Figure 10 A) at 5 ' non-translational region.
Pi72 constitutive gene encoded protein sequences that gene comprises are shown in Pi7 SEQ ID No.3 in the sequence table and Pi7SEQ ID No. 4.
Pi7-11 protein polypeptide of forming by 1142 amino-acid residues of genes encoding, molecular weight is 126.60 KD, iso-electric point is 6.46.Utilize this protein polypeptide of COIL analysis revealed to have CC (coiled-coil) structural domain.Pi7-1 albumen belongs to NBS-LRR albumen, and conservative kinase 1a (GLPGGGKTTVAR) is positioned at the 290th amino-acid residue of this polypeptide in the NBS structural domain; Kinase 2 (NKKYLIVIDDIW) is positioned at the 376th amino-acid residue of this polypeptide; Kinase 3a (DLGGRIIMTTRLNSI) is positioned at the 402nd amino-acid residue of this polypeptide, and GLPL (EDSPCYDIVNMCYGMPLALIW) is positioned at 461 amino-acid residues of this protein polypeptide.And 610-960 amino-acid residue of this proteic C-end is 16 imperfect LRR repetitions, and its leucine content is 14.0%, and its C-terminal also has the aminoacid sequence (Fig. 9 B) of a non-LRR structure (CNL).Two aminoacid sequences that indicate that roll off the production line do among the figure
Pi7The amino acid replacement (substitution) of gene specific, the gene specific acidic amino acid replacement site of this site and following another constitutive gene combines, and can differentiate
Pi7Gene and other allelotrope (
Pik,
Pik-m,
Pik-p,
Pik-s) difference (seeing embodiment 9 for details).
Pi7-21 protein polypeptide of forming by 1021 amino-acid residues of genes encoding, molecular weight is 114.57 KD, iso-electric point is 8.64.Utilize this protein polypeptide of COIL analysis revealed not have the CC structural domain, find to have the CC structural domain but utilize more powerful forecasting tool Paircoil2 to analyze.Pi7-2 albumen belongs to NBS-LRR albumen, and conservative kinase 1a (GFGGVGKTTIA) is positioned at the 211st amino-acid residue of this polypeptide in the NBS structural domain; Kinase 2 (KSYILLIDDIW) is positioned at the 330th amino-acid residue of this polypeptide; Kinase 3a (GGRIIVTTRFQAV) is positioned at the 358th amino-acid residue of this polypeptide; GLPL (EQVPEEIWKICGGLPLAIV) is positioned at the 415th amino-acid residue of this polypeptide; RNBS-D (CLLYLSIFPKGWK) is positioned at the 488th amino-acid residue of this polypeptide, and MHDV (KTFQVHDMVLEYI) is positioned at the 553rd amino-acid residue of this polypeptide.And 610-960 terminal amino-acid residue of this proteic C-is 13 imperfect LRR repetitions, and its leucine content is 17.0% (Figure 10 B).Two aminoacid sequences that indicate that roll off the production line do among the figure
Pi7The amino acid replacement (substitution) of gene specific, the gene specific acidic amino acid replacement site of this site and above-mentioned another constitutive gene combines, and can differentiate
Pi7Gene and other allelotrope (
Pik,
Pik-m,
Pik-p,
Pik-s) difference (seeing embodiment 9 for details).
Utilize the quantitative RT-PCR technology right
Pi7The expression of gene pattern is analyzed.The different time point in disease-resistant variety IRBL7-M inoculation back (0 hr, 12 hr, 24 hr, 72 hr 120h) go up and gather blade and extract its total RNA, utilize reverse transcription test kit SuperScript
TMReverse Transcriptase II carries out the synthetic of reverse transcription cDNA article one chain.The primer of RT-PCR does,
RRT5F：GAAGCTCTGATCAACGGTATTCC，
RRT5R：TCTTTGATCATCTTCGGGATACG；
RRT17F：TGAACTTCCACGATTGGATCCAC，
RRT17R：ACATGGTTCTTGAATACATCATGTC 。
Quantitative RT-PCR uses CFX96 Real-time PCR detection system and SYBR Premix Ex Taq test kit (precious biotech firm), and operation is carried out according to the test kit explanation.Figure 11 (left side) is a constitutive gene
Pi7-1Relative expression's level at different inoculation time points this shows, this expression of gene type is progressively to raise from 12 hr-72 hr, then progressively downward modulation.Figure 11 (right side) is another constitutive gene
Pi7-2Relative expression's level at different inoculation time points this shows, this expression of gene type is that 12 hr-24 hr progressively raises, then progressively downward modulation.It can also be seen that thus this gene is at inoculation contrast (H
2O) and between the inoculation pathogenic bacteria show visibly different inducible expression.2 genes all show as constitutive expression because the inoculation before with can both detect 2 expression of gene afterwards.
Of embodiment 7,
Pi7The amino acid replacement site that has a gene specific in 2 constitutive genes of gene respectively; Utilize these 2 sites to develop the molecule marker Pi7-SNP-1 (Pi7-SNP-1F:GCAGGTCAGCCAAGCAAT of gene specific respectively; Pi7-SNP-1R:CAACCGTTGTTTTGCCTCC, amplified production is used restriction enzyme
MseI carries out enzyme and cuts) and Pi7-SNP-2 (amplified production is used restriction enzyme for Pi7-SNP-2F:CGTGGAAGTTCAACAAAAGG, Pi7-SNP-2R:CAGCACCTGTATTATCCCAT
NdeI carries out enzyme and cuts), the two combines both to have can be used for differentiating
Pi7Gene and other allelotrope (
Pik,
Pik-p,
Pik-m,
Pik-s) difference (enzyme of amplified production is cut the effect difference; The swimming lane 1 of Figure 12 compares with swimming lane 2-5), also can be used for identifying filial generation
Pi7Range gene type on the site, promptly
Pi7/Pi7With
Pi7/pi7Plant (enzyme is cut the effect difference; The swimming lane 1 of Figure 12 compares with swimming lane 6-7).These information can be applied in the molecular marker assisted selection breeding process, to improve the purpose and the efficient of breeding.
Will
Pi7Gene clone is to plant conversion carriers such as pCAMBIA1300; In the agrobacterium strains such as importing EHA105; Be used to transform susceptible rice varieties Longjing 24 (imperial round-grained rice 24) and Q1063, obtained 285 strain transformed plants altogether, wherein 155 strains performance resistance reactions (table 1).This explanation can be used
Pi7Gene transformation paddy rice susceptible variety after a series of procedures of breeding such as selfing, purifying selection, can produce disease-resistant variety and in being applied to produce.
The sequence explanation
SEQ ID NO. 1 & 2 is
Pi7-1With
Pi7-2Nucleotide sequence; SEQ ID NO. 3 & 4 are coded products of SEQ ID NO. 1 & 2; SEQ ID NO. 5 & 6 have promotor
Pi7-1With
Pi7-2Nucleotide sequence.SEQ ID NO. 7 & 8 are used for amplification
Pi7-12Primer right; SEQ ID NO. 9 & 10 are respectively that primer is to RRT5F, RRT5R and RRT17F, RRT17R with SEQ ID NO. 11 & 12; SEQ ID NO. 13 & 14 are that primer is to Pi7-SNP-1F and Pi7-SNP-1R; SEQ ID NO. 15 & 16 are that primer is to Pi7-SNP-2F and Pi7-SNP-2R; SEQ ID NO. 17 ~ 26 is respectively
Pi7-1Kinase 1a, kinase 2, kinase 3a, GLPL sequence and
Pi7-2Kinase 1a, kinase 2, kinase 3a, GLPL, RNBS-D, MHD sequence.
Sequence table
< 110>Agricultural University Of South China
< 120>rice blast resistance gene Pi7 and application thereof
<130> *********
<160> 22
<170> PatentIn version 3.5
<210> 1
<211> 9101
<212> DNA
< 213>Oryza paddy rice (Orysa sativa L.)
<220>
<221> 5'UTR
<222> (1)..(104)
<220>
<221> CDS
<222> (105)..(668), (819)..(1387), (4160)..(6455)
<220>
<221> 3'UTR
<222> (6456)..(6645)
<400> 1
aaccttgctc taattacact cggagtcgga gagtcaggag gtgagagagt agcactcaac 60
gcaaaagggg atcggccgag ccgccgatcg aggcgcgagt agggatggag gcggctgcca 120
tggccgtaac cgcagccacg ggggccttgg cgcccgtgct agtgaagctg gccgctttgc 180
tggacgacgg ggagtgcaat cttctggagg ggagccggag cgacgcagag ttcatcagat 240
ccgagctgga ggccgttcat tctctcctca ccccaaatat cttggggagg atgggggatg 300
acgatgcggc gtgcaaggat ggcttgattg cggaggtccg ggagctgtcc tacgacctgg 360
atgatgccgt cgacgacttc ttggagctca atttcgagca gcgaagaagc gcaagccctt 420
tcggtgagct caaggcaaga gttgaggagc atgtctccaa tcgcttctct gactggaagc 480
taccggcggc gagccttccg ccgtcgtcgg tacaccgccg agctggcttg ccgccaccag 540
atgcagagct ggtggggatg gacaaacgta tggaagagct caccaaattg ctggaacaag 600
ggagcaatga tgcttcacga tggcgcaagc gaaaaccgca tttcccgctc agaaaaacag 660
ggctaaaggt acggttggat ctcgattctt cacaaatata gcttcctccg atagcagtgc 720
caatgaattt gtttacttct ctcttgattc tttaatttgg aagtactgta taacaaacat 780
ggagggatcg tcatttaact taattttttt tgttgcagca aaaaatcgtg atcaaggttg 840
ccatggaggg caataattgc cgttcaaaag caatggcttt agttgcgagc actggaggag 900
tggactcggt tgcgctcgta ggtgatctaa gagacaagat agaggtggtc ggttatggca 960
ttgaccccat caagctgatc tccgcgctcc ggaagaaggt gggcgatgcg gagttgctgc 1020
aggtcagcca agcaaacaaa gatgtgaagg agacgacgcc gatgcttgcg ccggtgaaat 1080
ccatatgtga atttcacaag gtcaaaacag tttgcatcct tggattgcca ggtggaggca 1140
aaacaacggt tgccagagaa ttatatgacg ccttgggaac gcacttccca tgccgggttt 1200
tcgtgtcagt ctctccaagt tctagtccca gtcccaatct cacaaagact cttgcagaca 1260
ttttcgctca agcacaacta ggagtaaccg atacacttag cacaccatat ggtgggagtg 1320
ggaccgggag agctcttcaa caacatctca tcgacaacat atcagctttc ctcctcaaca 1380
aaaagtaagc aatatcttat atctcttatc tagcttgtcc ttaatccggc ttcccaaatt 1440
aaagtgaaca ggctatatgt ccatataatg ttgggttttt ttttttctgt cctcgatgag 1500
gtatatatca agatatgcca aacttttttt aattagcgtt actttattct acctagtaaa 1560
tagatacaga aatggggcag gcccctcctt tgtgacaggc caatggtttt atgatgtgga 1620
aacaactccc ttatcactga agctcttgct attactcgtg atggtactga tcttgcaaag 1680
gcttgcagta tttgtaaatc cacagggagt ggtcatcttt ttttttttaa gataatgaat 1740
agaaatctgg cctctatata gaaagccaaa ggttacacaa gtgcatacac tccaaatctc 1800
aaagctgaga aacacgaaaa acaaacgacc aaaaagaaaa gacgataaga ttaagtttca 1860
agcttttcct ccttgcttca gaaaacaggg aaatgcagag ctagcccgct cttcccctgc 1920
catgccctct cactgccatt gcaactacca caaaagctcc cctgtttcgc cgtgagcgaa 1980
gggaggggat ggaactagtc tgctgctttg ttgtcgccaa gcactggttt catgctgccc 2040
aaacaaccta tgccccgcta gcgataacga gtctgttgcc gccatgctgt tgtcgctgga 2100
gagcacttgt cgctgctcag ctatgtgtta agtaagctac taacaatctc tccattctct 2160
tgtagactgc ttataaaaga ttgtacttca actttttatt aatgaaatag agttcctgcc 2220
ttacaacaac aagaacgaga gagagagaga gagccaatat cacgagttag cactgtgaag 2280
acttcctgac tgttatgtct tcaagaacag tataaaagca tttgccacta taaaaatatg 2340
atagccctca agatcattaa aaatatgtca tgttctttta ccttcataac aaccagtatc 2400
acaaatatga aaacatttaa aggaaaatca gtatttgcca tttatgtaaa gaaaaactag 2460
actagaactc tgggctttta ttttagtgca cactgtatac atgtgtgttc tttgacgtat 2520
atacttgata ttttttcaaa aacttttggg ggcatatctg taacccatgc cccatgcatc 2580
caaagtagta gtgagacaac tttcgtatca atggacatgt ggcattaaat ttaatcccaa 2640
tatttgttct ctcgaagtcc tgaacatata attggacaaa attatataat ttaattaggc 2700
caattgagcc atatatacac acactataag aaacttctac aaaacaaaca ttcgctcctt 2760
ttaggttttg accagtctat acatgaacca gccaacggtg tgctggtctg actataaatg 2820
tttttttaga gagtaaaggc accttccagt acctctctcg aggacagtaa aaacacccat 2880
gctttaaggt tatgaacaac aaagcataga acaaccacaa gaaaaaaaaa aacaactaga 2940
acaagttctg gaactagtta gggattcggg aagccattgc ttgctggctt gggacaagct 3000
atgcctgcca ttggactatg gtggagactg ggcataaagg atctcaatct cttggagatt 3060
ggcctatgtc ctaggtggcc atggctgact gcaaggagat atatttcttg caacggaaat 3120
tttcattgca aaagctgaaa atcgttgcga tatgtaccta ttgtgacata attgaatcca 3180
ttgggagccg tagcaataaa tcccgtggta ataggctatt gcaatgattt tccgatggtt 3240
gcataaaaag gtgctattac aatggttttg cattgctatt gcaacaactt gatatgtcat 3300
aggtatttat tgcaacagtt tgtaccatgc tgcaataaga tttcgcaaca cagacctatt 3360
cattaaatag cctatttcca cacattcatt tttgtggcaa taattcagtg cgacaagcat 3420
gttctatagc tataacctga tgcaacgggt tagacttgtt gcagtaactt attagcacat 3480
ttaatttttt actaaattta tattgattca actatttcaa catcattcaa tacagtaaat 3540
actggaaaat gcaagcaatc tccaaaaatt caccaataag agaaacgaat taaatcatat 3600
acacaattat ttcacaccgt acagtgcact taagtgtagg aaatataatg ctatatacat 3660
ttccaaccct caatacaggt tctgtaaata atgaaaagaa acatcaattt gatttcttga 3720
ccggtccctc tgagacacac agtgctttcc tcagcaacat cgttgccgat ctatgtaact 3780
tgtaaactcc ttgtgcttaa taaattctgg ccagggttac ttcagcctgc atggcgaaaa 3840
aaaagaactt gttagggaga tcgcaaacaa atgttagctg ccccatttct tctctactga 3900
ccataaaagc tctaggatca ggtcatgaaa acaatgccgt cagaccggaa atactcaaga 3960
ttcagaacac gactcctagg aactgcaagt ttaaatttca acaaaagcaa aatacaacca 4020
gatgacaaac taatatatat cagctgactg ttcatctcaa gtggactcac atctgtgaaa 4080
ttgttagttc tatgcatata tgctaatgta atactaatat gctattgagg cttattactc 4140
ttctcgactt ttataggtat ctcattgtaa tcgatgacat ttggcattgg gaagaatggg 4200
aagtcatcag aaagtccatt cccaagaatg atctgggtgg tagaataatc atgactactc 4260
gtcttaattc aatagctgag aagtgccaca ctgatgacaa tgatgttttt gtctacgaag 4320
ttggggatct agataataat gatgctttgt cgttgtcttg ggggatagca acaaagtctg 4380
gggcaggcaa caggatcgga actggagagg ataatccatg ctatgatatt gtgaacatgt 4440
gttatggtat gcctttagca cttatttggc tgtcgtcagc attggttgga gagatagaag 4500
aattaggtgg tgctgaagtg aaaaaatgta gggatttgag acacatagag gatggtattt 4560
tggacatccc atccttacaa ccattggcgg agagtttatg ccttggttat aaccatcttc 4620
ctctttatct gaggactttg ttgttgtact gtagtgcata ccattggtct aacagaatcg 4680
aaaggggtcg tctggtcagg aggtggattg cggaaggatt tgtgtcggaa gagaaagaag 4740
cagaaggtta ctttggcgag cttattaaca gaggatggat tacgcagcac ggagacaaca 4800
acagttataa ttactatgag atccaccccg tgatgctggc cttcctgaga tgcaagtcca 4860
aggagtacaa ttttttaaca tgcttgggtc tgggatctga tactagtact agtgcatcct 4920
ccccaaggtt gattcgccgg ctgtctcttc agggggggta tccagtggac tgcttgtcaa 4980
gcatgagtat ggatgtgtca cacacttgca gccttgtcgt ccttggtgac gtggcgcgac 5040
ccaagggaat ccccttctat atgtttaagc gcttgcgagt gttggacctt gaagataata 5100
aggatataca ggattctcat ctgcagggca tatgtgaaca gttaagcctc agagtgaggt 5160
accttggtct caagggaacg cggatccgaa agctccctca ggagatgagg aagctgaagc 5220
atttggagat tttgtatgtg gggagcactc ggatcagtga acttccgcaa gagattggag 5280
agctgaagca tctgcggatt ctggacgtga gaaacacgga catcactgag ctcccactgc 5340
agatacggga gctgcagcat ctgcacactc tggacgtgag gaacactcca atcagtgagc 5400
tcccgccgca ggttggcaag ctgcagaatc tcaagattat gtgcgtgagg agcactgggg 5460
ttagggagct cccaaaggag attggggagc tgaatcatct acagactctg gacgtgagaa 5520
acacgagggt gagagagctg ccatggcaag ctggccagat ctcccaatcg ttgcgcgtgc 5580
ttgccggtga cagtggcgat ggcgtgcggt tgcccgaagg cgtctgcgaa gctctgatca 5640
acggtattcc aggggctacg cgtgcaaaat gcagggaggt tctgtccatc gcgatcatcg 5700
atcgtttcgg acctcccctt gttgggatat tcaaagttcc cggcagtcat atgcgtatcc 5760
cgaagatgat caaagaccac ttccgcgttc tttcttgcct agacatcagg ctctgccaca 5820
agcttgagga tgatgaccaa aagttcctcg ccgagatgcc caacctgcag acgctcgtgc 5880
tgaggttcga ggccctacca agacaaccca taaccatcaa cggcacaggc ttccagatgc 5940
tggagagctt ccgtgtcgac agccgggtgc caaggatagc cttccatgaa gacgccatgc 6000
ccaacctcaa gcttctcgag ttcaagttct acgccggccc agcaagcaac gatgccatcg 6060
gcatcaccaa cctgaagagc ctccaaaagg tggtctttcg gtgctcgcca tggtacaaga 6120
gcgacgcccc tggcatcagc gccaccattg acgtcgtgaa gaaagaagcc gaggagcatc 6180
ccaaccggcc gatcaccctc ctcatcaatg ctgggtataa ggagatatca actgagtcac 6240
acgggagcag tgaaaacatt gcgggcagca gtgggatcga tactgagcct gcacaggcac 6300
agcatgataa tctccctgct gttcgagatg actacaaggg aaaagggatt cttcttgatg 6360
gcaggtgtcc tacctgcggc cgagcgacta aaattgaaga ggaaacccaa gatcgagtag 6420
cagatattga aattcaaaca gaaactacta gctagctagt accgctgtct aattttattt 6480
gtattaaata ctctccttca tgtaatactc taagtccccg ctcaattttt cttgcctccg 6540
caactctaca tgatactcaa aactgatttc ttacctccct ttgattgtag aaaaaacatg 6600
ggaagtttgg aggatttcaa tcctatgaaa aggactttga gaaacaaatt acggaatcct 6660
atcttttcct ttaaaagata tataatagaa aatcctatag agatttttat aggaaagtta 6720
acaaaagcct caatcttatg gagaattcct gtgtttattt gcatagagga aagtgagact 6780
cacctaatca ggggcggttc tagtgggtgg tccaggtgat ccctggacca ccctaaaatt 6840
tggcccatga gaaccgcccc ccccctcctc ctcccggggt tggtcgaaac cagaaaaaag 6900
gccactgagc taagctaggg ttagggagag gtgtagccat gaactagcca cagcagcacg 6960
catcgctggc tgccgcctcc gcctctgcgt cagcgccagc gtgccgtctc cccggctctc 7020
gactctgcct ccgcctccgc cttggcgccg atgcgtcgcc tcctcggctc ccgcttctac 7080
ctcagcgcca gcgtgccgcc tccgccttag cgccgcctcc tcgcctcctc ccttccccga 7140
ctcctccagt cccccactcc tgtcggttgc tatcgtcggc gggcggcgac caacgcctac 7200
aagccaacta cggctgcctg ggcaagccgg cggccacccc cgcccctgga aagggaaaac 7260
attgccattc cacaggcggc gccaggtccc ctttctctct agctttcttg ttgctcagca 7320
aagttcaaag gtcagtagac atttcttgtt ggttttttgc aatgttgccc tatgcagtag 7380
tgagtagtca gtactcagta gtagtgctag tgaaaaattt ctgccttgag tgtagtgaaa 7440
agcaagctga gaaatagcat gtgtgacaaa ttattaaatg atggtttaat cacatctatt 7500
gagcgggtta ttttctctca agttagcaaa gaagacatta tgcataattt catgtctata 7560
tatgaacgga gagtagaaaa gaagtagggt tttgtaatct tctacttatc tattataaga 7620
tatttgaatg attatgttct aactttcgta acttcaaatt tagatgttgg ttttaagcaa 7680
gttgtgatgt tgtgagatat attgctacat ttcaattata tttttattat ttttgacaca 7740
tttggcttga atttgtaaca atgtgacaat ttggagcatc aagtaggact acccgacgat 7800
tcatgaaacc aatgtgccga tatttagcgg ttcttaatca tactgaggta agaaggcaac 7860
attttgcaaa agtagaattt cttcatgaaa atcaggctta tagctttaat gttcacaaac 7920
tagctaagtt tccttgtacc ttagaggagg ggcatcatgt tcgattctca aatactcctt 7980
gtaatctaag taggaatgta aataatttgg ttgttaatga ataaaatgcc agaattttat 8040
tctaaaagaa tatgtaattc aaattaatat tattctagat gtatcttgaa tagcccatta 8100
gcactggacc agtctttcaa gttgggtcag tggtcggtac cattgcctgg cccatcatgg 8160
aacagaatgg ccatggtctg atcgcaatgg gccgaggcat gacgcgctgc gctttcgaaa 8220
tcacgggcct gagctgtcat gagggctatc ctgggccagg cccgcgagac aagagggtgt 8280
cggacaatgc aacaacaacg agaggcaata agcagagctt atgcattgca cgaccaacaa 8340
agttgtcact tacatccctt attattctca atcttaatac tatctccgtt caaaaaaaag 8400
tagactttta gatacgaatg tgaacaaacg tcctcatcaa tcttgcctgc tgataaagca 8460
accgcagcta ccacgcacta tgcagggatc gatcgatcga tggatggaca tattattact 8520
caacaaagta gaggcattaa tccgcttgca gactaatcag attcaggcga cagcacgcgg 8580
ctgctgattc atcagatggc attgcttcaa attgttcgct ttcaccatgc cctgcaccat 8640
gcatcattta gatatttatt ttactgtact catagtaatt aaccactgaa aattcactct 8700
aggaaatcgt gtgttaactg ttaacctggt agattttgat catgaattgg ttgatgtcgt 8760
cgagcgatat gacgccctcg tacgcggcgt tgaaacggta gctgtcgtca tcgcgtccag 8820
acgccaccag cagcggcggc gcaggcggca gctgctccac gcagctgtac tcgcactcgt 8880
cgtggcactc gccgtcgccg ccgccgccgc cgccgccgtt ctccgccgtg atcgccacct 8940
gcagcgtcag gtgcacctcc agcgtcacga acacgcccac caccttcgcc ggcgacccca 9000
tcgccgtcgg ccgctgcatc ttgtagctcg acatccacga gaatatgtac gccgccatcg 9060
acgacgagac gccgacgatg ttctgcttcc tcaggctgtt c 9101
<210> 2
<211> 7630
<212> DNA
< 213>Oryza paddy rice (Orysa sativa L.)
<220>
<221> 5'UTR
<222> (1)..(1232)
<220>
<221> CDS
<222> (1233)..(2224), (2389)..(4462)
<220>
<221> 3'UTR
<222> (4463)..(4706)
<400> 2
ggcctccgcc caaaagcctc acccccaagg ccccaatctt cccaccgcgc cgctgtcccc 60
cccccccccc ccccccccaa tcttctggcc tggccagctc tggtcggagg agggttcagt 120
tccatgggtg cgccccctgc ccatccctcc acttctactt tcgtcttgag atctcgccgt 180
ttctgatctc cttgccatgg caaccagttg atttggctgc caagatcgtc cgtgctttgc 240
aaaggtgaac aatctttttc ttcttgggct ttcactttgt tgtgttaata gactcagcgc 300
cgtcgcaact tgcaaataaa aaggttcaga tcggaaaatg gaaataccgt acacagtagt 360
ctatagagaa tcgactagta tttcctgact gtaagtaagg aagttgtagg taccatgttc 420
tgatgattat ctttgaagct tggaggggga caaaaagttg ggagagaaac aagaatgtat 480
ttgttcactt gattgggggc agtaggtaaa atctcagcgt taggaatttg cacctcttta 540
ttaagtttga ctaaatttat agaaaaaaat tagcaacact taaaacacca aattagtttc 600
attgaatcca acattgaata tattttgata atatgtttac tttgtgttca aaatgttact 660
atatttttct ataaatttga tcaaatttca ataagtttga ctagaaaaaa aagtcaaacc 720
gacttataat atgaatataa atggagggag tagattataa attatcttta ttataatact 780
ttgagtgtac aaacgaatga ataaagttta caaagttgaa aagttatttt taaatactat 840
tccgataaac ctgtatatag agaacttgtg aaaaaaaaca ccgtttgaaa tgtgtctgtg 900
ataatctata agaggaacgg ggcctaaata tattcctttt ttcctacgaa aatgcaagaa 960
gtattgccta atatattgat agagctgaag ttttaaagta aaatgaaaca tttgcatgag 1020
gcagcactgg acaaaaagcg cttgtgaaaa aaatatgttc caaatattat taattcattc 1080
atggtatggt ctttattttc attttccccc cttatccatg ggcaggacac ggaaattctc 1140
agcaggttcg cgcttgatct gaactgtctg tggctcatac tctcttgtgc ttgcaacagc 1200
tgaaagttgc agtgagaagt acagagaaca agatggagtt ggtggtaggt gcttccgaag 1260
ccaccatgaa atctctcttg ggcaagctgg gcaatcttct agcccaggag tatgctctca 1320
tcagcggtat ccgtggtgac atccagtaca tcaatgacga gcttgccagc atgcaggcct 1380
tcctccgtga tctcagcaac gtgccagagg gtcacagtca tggccaccgg atgaaggact 1440
ggatgaagca gatccgagac atcgcctatg atgttgagga ctgtatcgat gactttgccc 1500
accgcctccc tcaggattcc atcagcgatg ccaaatggtc cttcctactc acaaaaatct 1560
atgaactatg gacatggtgg ccacgtcgtg tgattgcttc caacattgcc caactcaagg 1620
tacgggcaca acagatcgca gatcgacgta gtagatacgg agtgaacaac ccagaacacc 1680
ttgacagtag cagcagtgcc aggacccgtg ctgtcaatta cgaaattgct gagtatcagg 1740
tcacaagccc tcagatcatt ggtataaagg agcctgtggg gatgaagacg gtcatggagg 1800
agcttgaggt ttggttaact aatcctcaag ctgaaaatgg gcaagctgtt ctgtccatag 1860
tcggttttgg aggtgtggga aagactacca ttgccacagc attgtacaga aaagtcagtg 1920
ataaatttca gtgccgggca tcagtagctg tgtctcagaa ctatgaccaa ggcaaagtcc 1980
tcaatagtat tctgagtcaa gtcagcaatc aggagcaggg cagcagcaca acaattagtg 2040
agaaaaagaa cctcacctca ggcgctaaga gcatgttgaa gacagccctg tcactgctca 2100
gaggtaattg tatatgtcag ccagaaaatg atggaaaccc tgataataca ccaatcaggc 2160
tgcaggaaac aacggacgat gatcaaaacc ccagaaaact ggaacagctc ctggccgaaa 2220
agaggtacct ttttttgtaa ataaaattgc tttgcttatc tgtaaattaa cttactcatc 2280
ccactctaaa tctaatgttt atttccttct atacacagca caactccatc ttttgaatgg 2340
gttttatttt tctcacttgt gctcattttt tttttatcat ctctgcagtt atatcctctt 2400
gattgatgac atttggtctg ccgaaacatg ggagagtatc agatcgattt tgcctaaaaa 2460
taataaaggc ggtagaataa tagtgactac aagatttcaa gctgttggtt caacatgctc 2520
ccctcttgaa actgatcgtt tgcatacagt tgattttctc accgatgacg agtcccaaaa 2580
cttattcaat acaagtattt gtgaatcaaa gataagaaaa gatagcaaca aagtagacga 2640
gcaagtccct gaggaaatat ggaaaatatg tgggggattg cctttggcca tagtcaccat 2700
ggctggtctt gtcgcctgca acccaaggaa agcctgctgc gattggagta aactttgcaa 2760
atcattattt ccagagcaag aaactcctct taccctcgat ggtgttacaa ggatactgga 2820
ttgttgttac aatgatttgc ctgcggatct gaagacttgc ttattgtact tgagtatatt 2880
tccgaagggt tggaaaatta gtaggaaacg tttgtcccgg cgatggatag ctgaaggttt 2940
tgctaatgag aagcaagggt taacccagga aagagttgca gaggcatact ttaatcaact 3000
cacaagaagg aacttagtac gtcccatgga gcatggcagc aatgggaagg taaaaacgtt 3060
tcaagttcat gacatggttc ttgaatacat catgtccaaa tcaatcgaag agaattttat 3120
tactgtggtt ggtggacact ggcagatgac tgcaccaagc aataaagtcc gtcgactgtc 3180
gatgcaaagc agtggatcca atcgtggaag ttcaacaaaa ggcctgaact tggctcaagt 3240
gagatcactg acggtgtttg ggaacctgaa ccatatgcca ttccattcat tcaactatgg 3300
gataatacag gtgctggatc ttgaggactg gaagggtttg aaagagagac atatgacgga 3360
gatatgtcaa atgcttttac tcaagtattt gagcatccga cgaacagaaa tttccaaaat 3420
tccctccaag attcagaaac ttgagtactt ggaaactctt gacataaggg agacatatgt 3480
cagggacctg cctaagtcaa tagtccagct aaaacggatc attagcatac ttggagggaa 3540
taaaaacaca cggaaggggc tgaggttgcc tcaagaaaaa agtaagaagc caattaaaaa 3600
cccgtcgcct caaggaaaaa caaaggagcc cgcaaagaaa ggattcttat cccaagaaaa 3660
aggtaaaggc gcaatgaaag cactccgtgt actgtcaggg attgagattg ttgaggaatc 3720
atcagaagta gctgcaggcc ttcatcagtt gacagggcta aggaagcttg ccatatacaa 3780
gctcaatata acaaagggtg gtgatacctt caaacaatta cagtcctcca ttgagtacct 3840
tggcagctgt ggtctgcaga ctctggccat caatgatgag aattctgaat ttatcaactc 3900
actgggcgac atgcccgcgc ctccaagata tcttgtcgcc cttgagctgt ctggcaagtt 3960
ggagaagcta cccaagtgga tcaccagcat cactactctc aacaagctaa ccatatctgt 4020
aacagttctt aggactgaaa ctttggagat cctccacatt ttaccttcat tgttttccct 4080
caccttcgcc ttttcactta gtgcagcgaa gcaggatcag gacataataa aggacatcct 4140
tgagaataat aaattggaca gtgatgggga aatcgtcatt ccagctgaag gattcaagag 4200
tcttaagctg cttcgcttct ttgcaccttt agtgccgaag ctcagctttt tggacaagaa 4260
tgcaatgcca gcactcgaaa tcattgaaat gcggtttaaa gacttcgaag gtctatttgg 4320
catcgaaatc cttgaaaatc tccgtgaggt gcatctcaaa gttagtgatg gggcagaagc 4380
aataaccaag ttccttgtaa atgatttgaa ggttaatact gagaaaccaa aagtatttgt 4440
tgatggcatc gtcactgcat gagaagtaaa attgctgcaa atcggagaac ttaccaatca 4500
tctgaggctt cccctctatt attactctct tagaatatat tgttattatt gctcaccttg 4560
caaaataaaa tagggatggc atagcatatt gctacaacgt accatggttc catcatagtt 4620
gatttcactt gtcattacag tgtctgttca gttgtgtttt ctattaataa aagggagatc 4680
tccgcaagaa accattatta tacttatatt cggttattga actctataaa tgatgggatt 4740
gctatattta tggtgccaca atttccatga gtgcggtatt tttttttcta gcagttggct 4800
ggtgttaaga tttgtgctgc cattgctcct ctattattgg tgcccaaaat tacgctttgc 4860
actatgttca cagttgtaaa acttacctaa atttcgtgta ttagtactgt aatgttgtga 4920
ttttcgcgtc cagttatatt tttttctttg ccagaagttt gatttcaagg tatatctggt 4980
aagcttcagc cggattcgtt aagtttttag tttaatcagc taaactttta acagtatgca 5040
tgcactagtt tctcccgtta tcttatccaa tataattaat gggctaattg gatctatgcc 5100
attacaaaca tggcaagatt cagaaaaatg ctactacaat ttgtctattc atagctgtgc 5160
tagtgaaatt ttacaaaatt ggaaccgtgc tattgatatc acgttttcca tccatctttt 5220
cttttttttc cttttcttct ttatcccgtc ttcttcccgc accgacgaga ggcgagtggc 5280
ggcgggaccg gcaagcgatg gcaggacctc ggcagctcag tggtggcagc ggcaaaaggg 5340
aaaggggaag gggtaggaga gaacctggcc cgccgacgca agcatgttgg tcgtcgtctt 5400
ccccgaggag cttgagcagt ggcaatggta gaggaatcca aggaggccga ggaggttgag 5460
cggcactaat ggtagaggat gccgaggagg tcgagcatca gcgagcttgc cctgggtggg 5520
cgagtgaggg agagctctcc caagtgtctg cgaagctctc atcttcgttt tcctcatccc 5580
cactttgctc ctcttccact actgctgttg aagctagcgg ccctctctca ccagatctgg 5640
gcggagctca agcgaggcgc caccaccgac gatggagctt cgcgtggacc ccacagcctc 5700
acccgattgc catccctcct accctcatca ctccatctcc aagcccaaat cccacctccc 5760
ccaccggctc ctcctcctca gcccctgccg tggcagccga ggccaccgat gggccaaccc 5820
tcaacatcgt ctcgaagcag ctccgcgcac tgtggaagaa gcacaaccaa atcctccaga 5880
tggaggagtc gctcactggc gggaggaagc tgaacaagga gcaggaggag gtgctccgat 5940
ccaagcccat cgtcatcgcg ctcattgatg agctcgagcg gatgcgtgcc ctgctcgccg 6000
acgccctcgc cgaggagctc tcctcctgcc ccgccccttc cctatctgtc actccctcct 6060
cctccacctc ctccggcgct gatttgtccg tcaaggatct cctcacgctc atctacttcg 6120
gctccctctt cgacgtcaag tcgcagatcg agttcgtcac caccatgctc gcacgctaag 6180
gagctagact gatgcatcac ctacgactat gtccgtcctt gccgcagcac gacttgctac 6240
ggtcccgcct cacgccccgt cgctcgccgg tcccgccgcc acttgcttct catcggtgcg 6300
ggaagaagat gggagaaaga aggaagaaaa cgggaaaaaa agagatggat gaaaaacgtg 6360
acagcaatag cacggttcca atttcgtaaa atttcaatgg caccgctacg aatagaagaa 6420
ttgtagtagc atttttctga atcaacatat ttgtaatggc atggatccaa taaaccccaa 6480
atttatttta tgccaataca aatattggca actggtgtca ttaacgtata aatgggtcgc 6540
ctacataaaa ccttccgcgt ccttcttggt tgaacaggta attaattgca accattttct 6600
gaaaaaattg actggaagtc tgaaaccgtc cctgctactc cctgaatgct ttagttcaat 6660
ctctatgaag ctccatccaa acaaaatctc cagatgatat ttcattaaaa acatcatgtc 6720
cagaactgtc cagtatccaa gaatggctat gatcttttct gaattctgga ctcgggactt 6780
atattacatg tatcaaatcc agttattttc tggagctaca ccagatgaaa tcaccaagta 6840
aaacaattat ataaggatgg tgagttgtta attaattaca gctttgatag accactctgt 6900
ttgttaatgc ctgacaagag ttcacaaaga aaaaaataca caggattaca aaacttttta 6960
gatactatct gatgtgtgtt aattctctca aatactatgt acctgaacaa acaagagaga 7020
acatgttgtg aattgtgatc tcctcatgcg agtgacctga aactctctac agggtcagac 7080
tcctcaagca tcactgcagc ctggaaacaa tcagcagcat cattgatcct cccatcattc 7140
ctgtgaactc tccctaaatg aagccaagcc atcctgttcg taggctcaat tctcagtgca 7200
tcagagagga agcatctcgc tgcagggaga taccttgacc cttgcttgca gagaagtgca 7260
ccaatggcca ccttggatgg aacatgctct agctctatcg agaatgcgtt gacgtaggca 7320
gccagtgcct ccttgttctg atctcgcgcc tcgagcatga aacctgaact tatagcatcc 7380
acgagtcaga ttttgatgtt gaattcagtt gaatgcagag aattgtcatg ttagtgttac 7440
atagaagtgg tgaaagttca agtttagttg agttgatcac cttctgcatg cattgttgca 7500
gccgagtagg attttagggc tctagccttc tgcagacata tctcagcgtc tctccagatt 7560
gagagactgg agtacaggtt tgcaagtcct tgccatattt caaactcact tacactgtca 7620
ttctgtccct 7630
<210> 3
<211> 1142
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 3
Met Glu Ala Ala Ala Met Ala Val Thr Ala Ala Thr Gly Ala Leu Ala
1 5 10 15
Pro Val Leu Val Lys Leu Ala Ala Leu Leu Asp Asp Gly Glu Cys Asn
20 25 30
Leu Leu Glu Gly Ser Arg Ser Asp Ala Glu Phe Ile Arg Ser Glu Leu
35 40 45
Glu Ala Val His Ser Leu Leu Thr Pro Asn Ile Leu Gly Arg Met Gly
50 55 60
Asp Asp Asp Ala Ala Cys Lys Asp Gly Leu Ile Ala Glu Val Arg Glu
65 70 75 80
Leu Ser Tyr Asp Leu Asp Asp Ala Val Asp Asp Phe Leu Glu Leu Asn
85 90 95
Phe Glu Gln Arg Arg Ser Ala Ser Pro Phe Gly Glu Leu Lys Ala Arg
100 105 110
Val Glu Glu His Val Ser Asn Arg Phe Ser Asp Trp Lys Leu Pro Ala
115 120 125
Ala Ser Leu Pro Pro Ser Ser Val His Arg Arg Ala Gly Leu Pro Pro
130 135 140
Pro Asp Ala Glu Leu Val Gly Met Asp Lys Arg Met Glu Glu Leu Thr
145 150 155 160
Lys Leu Leu Glu Gln Gly Ser Asn Asp Ala Ser Arg Trp Arg Lys Arg
165 170 175
Lys Pro His Phe Pro Leu Arg Lys Thr Gly Leu Lys Gln Lys Ile Val
180 185 190
Ile Lys Val Ala Met Glu Gly Asn Asn Cys Arg Ser Lys Ala Met Ala
195 200 205
Leu Val Ala Ser Thr Gly Gly Val Asp Ser Val Ala Leu Val Gly Asp
210 215 220
Leu Arg Asp Lys Ile Glu Val Val Gly Tyr Gly Ile Asp Pro Ile Lys
225 230 235 240
Leu Ile Ser Ala Leu Arg Lys Lys Val Gly Asp Ala Glu Leu Leu Gln
245 250 255
Val Ser Gln Ala Lys Lys Asp Val Lys Glu Thr Thr Pro Met Leu Ala
260 265 270
Pro Val Lys Ser Ile Cys Glu Phe His Lys Val Lys Thr Val Cys Ile
275 280 285
Leu Gly Leu Pro Gly Gly Gly Lys Thr Thr Val Ala Arg Glu Leu Tyr
290 295 300
Asp Ala Leu Gly Thr His Phe Pro Cys Arg Val Phe Val Ser Val Ser
305 310 315 320
Pro Ser Ser Ser Pro Ser Pro Asn Leu Thr Lys Thr Leu Ala Asp Ile
325 330 335
Phe Ala Gln Ala Gln Leu Gly Val Thr Asp Thr Leu Ser Thr Ser Tyr
340 345 350
Gly Gly Ser Gly Thr Gly Arg Ala Leu Gln Gln His Leu Ile Asp Asn
355 360 365
Ile Ser Ala Phe Leu Leu Asn Lys Lys Tyr Leu Ile Val Ile Asp Asp
370 375 380
Ile Trp His Trp Glu Glu Trp Glu Val Ile Arg Lys Ser Ile Pro Lys
385 390 395 400
Asn Asp Leu Gly Gly Arg Ile Ile Met Thr Thr Arg Leu Asn Ser Ile
405 410 415
Ala Gly Lys Cys His Thr Asp Asp Asn Asp Val Phe Val Tyr Glu Val
420 425 430
Gly Asp Leu Asp Asn Asn Asp Ala Leu Ser Leu Ser Trp Gly Ile Ala
435 440 445
Thr Lys Ser Gly Ala Gly Asn Arg Ile Gly Thr Gly Glu Asp Ser Pro
450 455 460
Cys Tyr Asp Ile Val Asn Met Cys Tyr Gly Met Pro Leu Ala Leu Ile
465 470 475 480
Trp Leu Ser Ser Ala Leu Val Gly Glu Ile Glu Glu Leu Gly Gly Ala
485 490 495
Glu Val Lys Lys Cys Arg Asp Leu Arg His Ile Glu Asp Gly Ile Leu
500 505 510
Asp Ile Pro Ser Leu Gln Pro Leu Ala Glu Ser Leu Cys Leu Gly Tyr
515 520 525
Asn His Leu Pro Leu Tyr Leu Arg Thr Leu Leu Leu Tyr Cys Ser Ala
530 535 540
Tyr His Trp Ser Asn Arg Ile Glu Arg Gly Arg Leu Val Arg Arg Trp
545 550 555 560
Ile Ala Glu Gly Phe Val Ser Glu Glu Lys Glu Ala Glu Gly Tyr Phe
565 570 575
Gly Glu Leu Ile Asn Arg Gly Trp Ile Thr Gln His Gly Asp Asn Asn
580 585 590
Ser Tyr Asn Tyr Tyr Glu Ile His Pro Val Met Leu Ala Phe Leu Arg
595 600 605
Cys Lys Ser Lys Glu Tyr Asn Phe Leu Thr Cys Leu Gly Leu Gly Ser
610 615 620
Asp Thr Ser Thr Ser Ala Ser Ser Pro Arg Leu Ile Arg Arg Leu Ser
625 630 635 640
Leu Gln Gly Gly Tyr Pro Val Asp Cys Leu Ser Ser Met Ser Met Asp
645 650 655
Val Ser His Thr Cys Ser Leu Val Val Leu Gly Asp Val Ala Arg Pro
660 665 670
Lys Gly Ile Pro Phe Tyr Met Phe Lys Arg Leu Arg Val Leu Asp Leu
675 680 685
Glu Asp Asn Lys Asp Ile Gln Asp Ser His Leu Gln Gly Ile Cys Glu
690 695 700
Gln Leu Ser Leu Arg Val Arg Tyr Leu Gly Leu Lys Gly Thr Arg Ile
705 710 715 720
Arg Lys Leu Pro Gln Glu Met Arg Lys Leu Lys His Leu Glu Ile Leu
725 730 735
Tyr Val Gly Ser Thr Arg Ile Ser Glu Leu Pro Gln Glu Ile Gly Glu
740 745 750
Leu Lys His Leu Arg Ile Leu Asp Val Arg Asn Thr Asp Ile Thr Glu
755 760 765
Leu Pro Leu Gln Ile Arg Glu Leu Gln His Leu His Thr Leu Asp Val
770 775 780
Arg Asn Thr Pro Ile Ser Glu Leu Pro Pro Gln Val Gly Lys Leu Gln
785 790 795 800
Asn Leu Lys Ile Met Cys Val Arg Ser Thr Gly Val Arg Glu Leu Pro
805 810 815
Lys Glu Ile Gly Glu Leu Asn His Leu Gln Thr Leu Asp Val Arg Asn
820 825 830
Thr Arg Val Arg Glu Leu Pro Trp Gln Ala Gly Gln Ile Ser Gln Ser
835 840 845
Leu Arg Val Leu Ala Gly Asp Ser Gly Asp Gly Val Arg Leu Pro Glu
850 855 860
Gly Val Cys Glu Ala Leu Ile Asn Gly Ile Pro Gly Ala Thr Arg Ala
865 870 875 880
Lys Cys Arg Glu Val Leu Ser Ile Ala Ile Ile Asp Arg Phe Gly Pro
885 890 895
Pro Leu Val Gly Ile Phe Lys Val Pro Gly Ser His Met Arg Ile Pro
900 905 910
Lys Met Ile Lys Asp His Phe Arg Val Leu Ser Cys Leu Asp Ile Arg
915 920 925
Leu Cys His Lys Leu Glu Asp Asp Asp Gln Lys Phe Leu Ala Glu Met
930 935 940
Pro Asn Leu Gln Thr Leu Val Leu Arg Phe Glu Ala Leu Pro Arg Gln
945 950 955 960
Pro Ile Thr Ile Asn Gly Thr Gly Phe Gln Met Leu Glu Ser Phe Arg
965 970 975
Val Asp Ser Arg Val Pro Arg Ile Ala Phe His Glu Asp Ala Met Pro
980 985 990
Asn Leu Lys Leu Leu Glu Phe Lys Phe Tyr Ala Gly Pro Ala Ser Asn
995 1000 1005
Asp Ala Ile Gly Ile Thr Asn Leu Lys Ser Leu Gln Lys Val Val
1010 1015 1020
Phe Arg Cys Ser Pro Trp Tyr Lys Ser Asp Ala Pro Gly Ile Ser
1025 1030 1035
Ala Thr Ile Asp Val Val Lys Lys Glu Ala Glu Glu His Pro Asn
1040 1045 1050
Arg Pro Ile Thr Leu Leu Ile Asn Ala Gly Tyr Lys Glu Ile Ser
1055 1060 1065
Thr Glu Ser His Gly Ser Ser Glu Asn Ile Ala Gly Ser Ser Gly
1070 1075 1080
Ile Asp Thr Glu Pro Ala Gln Ala Gln His Asp Asn Leu Pro Ala
1085 1090 1095
Val Arg Asp Asp Tyr Lys Gly Lys Gly Ile Leu Leu Asp Gly Arg
1100 1105 1110
Cys Pro Thr Cys Gly Arg Ala Thr Lys Ile Glu Glu Glu Thr Gln
1115 1120 1125
Asp Arg Val Ala Asp Ile Glu Ile Gln Thr Glu Thr Thr Ser
1130 1135 1140
<210> 4
<211> 1021
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 4
Met Glu Leu Val Val Gly Ala Ser Glu Ala Thr Met Lys Ser Leu Leu
1 5 10 15
Gly Lys Leu Gly Asn Leu Leu Ala Gln Glu Tyr Ala Leu Ile Ser Gly
20 25 30
Ile Arg Gly Asp Ile Gln Tyr Ile Asn Asp Glu Leu Ala Ser Met Gln
35 40 45
Ala Phe Leu Arg Asp Leu Ser Asn Val Pro Glu Gly His Ser His Gly
50 55 60
His Arg Met Lys Asp Trp Met Lys Gln Ile Arg Asp Ile Ala Tyr Asp
65 70 75 80
Val Glu Asp Cys Ile Asp Asp Phe Ala His Arg Leu Pro Gln Asp Ser
85 90 95
Ile Ser Asp Ala Lys Trp Ser Phe Leu Leu Thr Lys Ile Tyr Glu Leu
100 105 110
Trp Thr Trp Trp Pro Arg Arg Val Ile Ala Ser Asn Ile Ala Gln Leu
115 120 125
Lys Val Arg Ala Gln Gln Ile Ala Asp Arg Arg Ser Arg Tyr Gly Val
130 135 140
Asn Asn Pro Glu His Leu Asp Ser Ser Ser Ser Ala Arg Thr Arg Ala
145 150 155 160
Val Asn Tyr Glu Ile Ala Glu Tyr Gln Val Thr Ser Pro Gln Ile Ile
165 170 175
Gly Ile Lys Glu Pro Val Gly Met Lys Thr Val Met Glu Glu Leu Glu
180 185 190
Val Trp Leu Thr Asn Pro Gln Ala Glu Asn Gly Gln Ala Val Leu Ser
195 200 205
Ile Val Gly Phe Gly Gly Val Gly Lys Thr Thr Ile Ala Thr Ala Leu
210 215 220
Tyr Arg Lys Val Ser Asp Lys Phe Gln Cys Arg Ala Ser Val Ala Val
225 230 235 240
Ser Gln Asn Tyr Asp Gln Gly Lys Val Leu Asn Ser Ile Leu Ser Gln
245 250 255
Val Ser Asn Gln Glu Gln Gly Ser Ser Thr Thr Ile Ser Glu Lys Lys
260 265 270
Asn Leu Thr Ser Gly Ala Lys Ser Met Leu Lys Thr Ala Leu Ser Leu
275 280 285
Leu Arg Gly Asn Cys Ile Cys Gln Pro Glu Asn Asp Gly Asn Pro Asp
290 295 300
Asn Thr Pro Ile Arg Leu Gln Glu Thr Thr Asp Asp Asp Gln Asn Pro
305 310 315 320
Arg Lys Leu Glu Gln Leu Leu Ala Glu Lys Ser Tyr Ile Leu Leu Ile
325 330 335
Asp Asp Ile Trp Ser Ala Glu Thr Trp Glu Ser Ile Arg Ser Ile Leu
340 345 350
Pro Lys Asn Asn Lys Gly Gly Arg Ile Ile Val Thr Thr Arg Phe Gln
355 360 365
Ala Val Gly Ser Thr Cys Ser Pro Leu Glu Thr Asp Arg Leu His Thr
370 375 380
Val Asp Phe Leu Thr Asp Asp Glu Ser Gln Asn Leu Phe Asn Thr Ser
385 390 395 400
Ile Cys Glu Ser Lys Ile Arg Lys Asp Ser Asn Lys Val Asp Glu Gln
405 410 415
Val Pro Glu Glu Ile Trp Lys Ile Cys Gly Gly Leu Pro Leu Ala Ile
420 425 430
Val Thr Met Ala Gly Leu Val Ala Cys Asn Pro Arg Lys Ala Cys Cys
435 440 445
Asp Trp Ser Lys Leu Cys Lys Ser Leu Phe Pro Glu Gln Glu Thr Pro
450 455 460
Leu Thr Leu Asp Gly Val Thr Arg Ile Leu Asp Cys Cys Tyr Asn Asp
465 470 475 480
Leu Pro Ala Asp Leu Lys Thr Cys Leu Leu Tyr Leu Ser Ile Phe Pro
485 490 495
Lys Gly Trp Lys Ile Ser Arg Lys Arg Leu Ser Arg Arg Trp Ile Ala
500 505 510
Glu Gly Phe Ala Asn Glu Lys Gln Gly Leu Thr Gln Glu Arg Val Ala
515 520 525
Glu Ala Tyr Phe Asn Gln Leu Thr Arg Arg Asn Leu Val Arg Pro Met
530 535 540
Glu His Gly Ser Asn Gly Lys Val Lys Thr Phe Gln Val His Asp Met
545 550 555 560
Val Leu Glu Tyr Ile Met Ser Lys Ser Ile Glu Glu Asn Phe Ile Thr
565 570 575
Val Val Gly Gly His Trp Gln Met Thr Ala Pro Ser Asn Lys Val Arg
580 585 590
Arg Leu Ser Met Gln Ser Ser Gly Ser Asn Arg Gly Ser Ser Thr Lys
595 600 605
Gly Leu Asn Leu Ala Gln Val Arg Ser Leu Thr Val Phe Gly Asn Leu
610 615 620
Asn His Met Pro Phe His Ser Phe Asn Tyr Gly Ile Ile Gln Val Leu
625 630 635 640
Asp Leu Glu Asp Trp Lys Gly Leu Lys Glu Arg His Met Thr Glu Ile
645 650 655
Cys Gln Met Leu Leu Leu Lys Tyr Leu Ser Ile Arg Arg Thr Glu Ile
660 665 670
Ser Lys Ile Pro Ser Lys Ile Gln Lys Leu Glu Tyr Leu Glu Thr Leu
675 680 685
Asp Ile Arg Glu Thr Tyr Val Arg Asp Leu Pro Lys Ser Ile Val Gln
690 695 700
Leu Lys Arg Ile Ile Ser Ile Leu Gly Gly Asn Lys Asn Thr Arg Lys
705 710 715 720
Gly Leu Arg Leu Pro Gln Glu Lys Ser Lys Lys Pro Ile Lys Asn Pro
725 730 735
Ser Pro Gln Gly Lys Thr Lys Glu Pro Ala Lys Lys Gly Phe Leu Ser
740 745 750
Gln Glu Lys Gly Lys Gly Ala Met Lys Ala Leu Arg Val Leu Ser Gly
755 760 765
Ile Glu Ile Val Glu Glu Ser Ser Glu Val Ala Ala Gly Leu His Gln
770 775 780
Leu Thr Gly Leu Arg Lys Leu Ala Ile Tyr Lys Leu Asn Ile Thr Lys
785 790 795 800
Gly Gly Asp Thr Phe Lys Gln Leu Gln Ser Ser Ile Glu Tyr Leu Gly
805 810 815
Ser Cys Gly Leu Gln Thr Leu Ala Ile Asn Asp Glu Asn Ser Glu Phe
820 825 830
Ile Asn Ser Leu Gly Asp Met Pro Ala Pro Pro Arg Tyr Leu Val Ala
835 840 845
Leu Glu Leu Ser Gly Lys Leu Glu Lys Leu Pro Lys Trp Ile Thr Ser
850 855 860
Ile Thr Thr Leu Asn Lys Leu Thr Ile Ser Val Thr Val Leu Arg Thr
865 870 875 880
Glu Thr Leu Glu Ile Leu His Ile Leu Pro Ser Leu Phe Ser Leu Thr
885 890 895
Phe Ala Phe Ser Leu Ser Ala Ala Lys Gln Asp Gln Asp Ile Ile Lys
900 905 910
Asp Ile Leu Glu Asn Asn Lys Leu Asp Ser Asp Gly Glu Ile Val Ile
915 920 925
Pro Ala Glu Gly Phe Lys Ser Leu Lys Leu Leu Arg Phe Phe Ala Pro
930 935 940
Leu Val Pro Lys Leu Ser Phe Leu Asp Lys Asn Ala Met Pro Ala Leu
945 950 955 960
Glu Ile Ile Glu Met Arg Phe Lys Asp Phe Glu Gly Leu Phe Gly Ile
965 970 975
Glu Ile Leu Glu Asn Leu Arg Glu Val His Leu Lys Val Ser Asp Gly
980 985 990
Ala Glu Ala Ile Thr Lys Phe Leu Val Asn Asp Leu Lys Val Asn Thr
995 1000 1005
Glu Lys Pro Lys Val Phe Val Asp Gly Ile Val Thr Ala
1010 1015 1020
<210> 5
<211> 10321
<212> DNA
< 213>Oryza paddy rice (Orysa sativa L.)
<220>
<221> promoter
<222> (1)..(1220)
<220>
<221> 5'UTR
<222> (1221)..(1324)
<220>
<221> CDS
<222> (1325)..(1888), (2039)..(2607), (5380)..(7675)
<220>
<221> 3'UTR
<222> (7676)..(7865)
<400> 5
gggggggaca gcggcgcggt gggaagattg gggccttggg ggtgaggctt ttgggcggag 60
gccaataata ctgcactgca ttcgcagact cgttgactga ctcgtggtca tgccgtttct 120
ccctaatatt tcctccgtcc tttaatataa tattaaagtc tttaatatta tacttgtact 180
actccctcca ttataaaata taagtcataa ccacccctaa cccaaacacc aagaaaataa 240
ttattattat cttgtagttt agaacatcct aataaataca atgcatgcat ccaatagaat 300
cagagagcat gaataataat ttaataaaaa agaggtcata gttaattgcc tatatgcatg 360
catgccttat attatgaaac atctaagaaa agtggttgtg ccttatatta tggaatagag 420
ggagtcctat ttaaccattt atcttatttt aaaaaaatta tgcaaataat aaaaataaaa 480
agttgtgctt aaaagatttt agataataaa ataagtaaaa gaaaattaaa taataatttt 540
caaacttttt tttaataaaa cgaaggatcg tactccctcc attctaaatt gatctacgta 600
tttcataggt acaccaagac caagaaaaac taataactct ttcatattat atttactcta 660
acaacaaact caatgcatgc atcatcccca atatttccta gccaatagca aatcaagata 720
ttgcatgtgg gttataaata cttgtgtgca tggaagcatg catcaatgtc catttactcc 780
aatgcacaca aataacgaat agacttaata aatgaacaca aatatataga tcatttagga 840
ataacctaaa aaaactatat atagatcaat ttagaatgaa gggagtacaa agtaaaatgt 900
taaaatacct tatattagag aacaaacgaa ctaattatgt gaaagcagtt ctctaaccca 960
ttgcgagagg accgagaaaa ataccaccac tttaagagca gttacaatag caggctataa 1020
gccaactgca aacatatttt aaggagataa ataaggagag agaagggtag cgggctatag 1080
atttgtagcc aactgtagca cagactccaa gacacagtgt gtatgacagg tgggaccagg 1140
tattaatagt atagtacttg tatgaataag ctattagatt ggctataaat aaattgaagc 1200
tactacttgg ctgtactgtt aaccttgctc taattacact cggagtcgga gagtcaggag 1260
gtgagagagt agcactcaac gcaaaagggg atcggccgag ccgccgatcg aggcgcgagt 1320
agggatggag gcggctgcca tggccgtaac cgcagccacg ggggccttgg cgcccgtgct 1380
agtgaagctg gccgctttgc tggacgacgg ggagtgcaat cttctggagg ggagccggag 1440
cgacgcagag ttcatcagat ccgagctgga ggccgttcat tctctcctca ccccaaatat 1500
cttggggagg atgggggatg acgatgcggc gtgcaaggat ggcttgattg cggaggtccg 1560
ggagctgtcc tacgacctgg atgatgccgt cgacgacttc ttggagctca atttcgagca 1620
gcgaagaagc gcaagccctt tcggtgagct caaggcaaga gttgaggagc atgtctccaa 1680
tcgcttctct gactggaagc taccggcggc gagccttccg ccgtcgtcgg tacaccgccg 1740
agctggcttg ccgccaccag atgcagagct ggtggggatg gacaaacgta tggaagagct 1800
caccaaattg ctggaacaag ggagcaatga tgcttcacga tggcgcaagc gaaaaccgca 1860
tttcccgctc agaaaaacag ggctaaaggt acggttggat ctcgattctt cacaaatata 1920
gcttcctccg atagcagtgc caatgaattt gtttacttct ctcttgattc tttaatttgg 1980
aagtactgta taacaaacat ggagggatcg tcatttaact taattttttt tgttgcagca 2040
aaaaatcgtg atcaaggttg ccatggaggg caataattgc cgttcaaaag caatggcttt 2100
agttgcgagc actggaggag tggactcggt tgcgctcgta ggtgatctaa gagacaagat 2160
agaggtggtc ggttatggca ttgaccccat caagctgatc tccgcgctcc ggaagaaggt 2220
gggcgatgcg gagttgctgc aggtcagcca agcaaaaaaa gatgtgaagg agacgacgcc 2280
gatgcttgcg ccggtgaaat ccatatgtga atttcacaag gtcaaaacag tttgcatcct 2340
tggattgcca ggtggaggca aaacaacggt tgccagagaa ttatatgacg ccttgggaac 2400
gcacttccca tgccgggttt tcgtgtcagt ctctccaagt tctagtccca gtcccaatct 2460
cacaaagact cttgcagaca ttttcgctca agcacaacta ggagtaaccg atacacttag 2520
cacaccatat ggtgggagtg ggaccgggag agctcttcaa caacatctca tcgacaacat 2580
atcagctttc ctcctcaaca aaaagtaagc aatatcttat atctcttatc tagcttgtcc 2640
ttaatccggc ttcccaaatt aaagtgaaca ggctatatgt ccatataatg ttgggttttt 2700
ttttttctgt cctcgatgag gtatatatca agatatgcca aacttttttt aattagcgtt 2760
actttattct acctagtaaa tagatacaga aatggggcag gcccctcctt tgtgacaggc 2820
caatggtttt atgatgtgga aacaactccc ttatcactga agctcttgct attactcgtg 2880
atggtactga tcttgcaaag gcttgcagta tttgtaaatc cacagggagt ggtcatcttt 2940
ttttttttaa gataatgaat agaaatctgg cctctatata gaaagccaaa ggttacacaa 3000
gtgcatacac tccaaatctc aaagctgaga aacacgaaaa acaaacgacc aaaaagaaaa 3060
gacgataaga ttaagtttca agcttttcct ccttgcttca gaaaacaggg aaatgcagag 3120
ctagcccgct cttcccctgc catgccctct cactgccatt gcaactacca caaaagctcc 3180
cctgtttcgc cgtgagcgaa gggaggggat ggaactagtc tgctgctttg ttgtcgccaa 3240
gcactggttt catgctgccc aaacaaccta tgccccgcta gcgataacga gtctgttgcc 3300
gccatgctgt tgtcgctgga gagcacttgt cgctgctcag ctatgtgtta agtaagctac 3360
taacaatctc tccattctct tgtagactgc ttataaaaga ttgtacttca actttttatt 3420
aatgaaatag agttcctgcc ttacaacaac aagaacgaga gagagagaga gagccaatat 3480
cacgagttag cactgtgaag acttcctgac tgttatgtct tcaagaacag tataaaagca 3540
tttgccacta taaaaatatg atagccctca agatcattaa aaatatgtca tgttctttta 3600
ccttcataac aaccagtatc acaaatatga aaacatttaa aggaaaatca gtatttgcca 3660
tttatgtaaa gaaaaactag actagaactc tgggctttta ttttagtgca cactgtatac 3720
atgtgtgttc tttgacgtat atacttgata ttttttcaaa aacttttggg ggcatatctg 3780
taacccatgc cccatgcatc caaagtagta gtgagacaac tttcgtatca atggacatgt 3840
ggcattaaat ttaatcccaa tatttgttct ctcgaagtcc tgaacatata attggacaaa 3900
attatataat ttaattaggc caattgagcc atatatacac acactataag aaacttctac 3960
aaaacaaaca ttcgctcctt ttaggttttg accagtctat acatgaacca gccaacggtg 4020
tgctggtctg actataaatg tttttttaga gagtaaaggc accttccagt acctctctcg 4080
aggacagtaa aaacacccat gctttaaggt tatgaacaac aaagcataga acaaccacaa 4140
gaaaaaaaaa aacaactaga acaagttctg gaactagtta gggattcggg aagccattgc 4200
ttgctggctt gggacaagct atgcctgcca ttggactatg gtggagactg ggcataaagg 4260
atctcaatct cttggagatt ggcctatgtc ctaggtggcc atggctgact gcaaggagat 4320
atatttcttg caacggaaat tttcattgca aaagctgaaa atcgttgcga tatgtaccta 4380
ttgtgacata attgaatcca ttgggagccg tagcaataaa tcccgtggta ataggctatt 4440
gcaatgattt tccgatggtt gcataaaaag gtgctattac aatggttttg cattgctatt 4500
gcaacaactt gatatgtcat aggtatttat tgcaacagtt tgtaccatgc tgcaataaga 4560
tttcgcaaca cagacctatt cattaaatag cctatttcca cacattcatt tttgtggcaa 4620
taattcagtg cgacaagcat gttctatagc tataacctga tgcaacgggt tagacttgtt 4680
gcagtaactt attagcacat ttaatttttt actaaattta tattgattca actatttcaa 4740
catcattcaa tacagtaaat actggaaaat gcaagcaatc tccaaaaatt caccaataag 4800
agaaacgaat taaatcatat acacaattat ttcacaccgt acagtgcact taagtgtagg 4860
aaatataatg ctatatacat ttccaaccct caatacaggt tctgtaaata atgaaaagaa 4920
acatcaattt gatttcttga ccggtccctc tgagacacac agtgctttcc tcagcaacat 4980
cgttgccgat ctatgtaact tgtaaactcc ttgtgcttaa taaattctgg ccagggttac 5040
ttcagcctgc atggcgaaaa aaaagaactt gttagggaga tcgcaaacaa atgttagctg 5100
ccccatttct tctctactga ccataaaagc tctaggatca ggtcatgaaa acaatgccgt 5160
cagaccggaa atactcaaga ttcagaacac gactcctagg aactgcaagt ttaaatttca 5220
acaaaagcaa aatacaacca gatgacaaac taatatatat cagctgactg ttcatctcaa 5280
gtggactcac atctgtgaaa ttgttagttc tatgcatata tgctaatgta atactaatat 5340
gctattgagg cttattactc ttctcgactt ttataggtat ctcattgtaa tcgatgacat 5400
ttggcattgg gaagaatggg aagtcatcag aaagtccatt cccaagaatg atctgggtgg 5460
tagaataatc atgactactc gtcttaattc aatagctgag aagtgccaca ctgatgacaa 5520
tgatgttttt gtctacgaag ttggggatct agataataat gatgctttgt cgttgtcttg 5580
ggggatagca acaaagtctg gggcaggcaa caggatcgga actggagagg ataatccatg 5640
ctatgatatt gtgaacatgt gttatggtat gcctttagca cttatttggc tgtcgtcagc 5700
attggttgga gagatagaag aattaggtgg tgctgaagtg aaaaaatgta gggatttgag 5760
acacatagag gatggtattt tggacatccc atccttacaa ccattggcgg agagtttatg 5820
ccttggttat aaccatcttc ctctttatct gaggactttg ttgttgtact gtagtgcata 5880
ccattggtct aacagaatcg aaaggggtcg tctggtcagg aggtggattg cggaaggatt 5940
tgtgtcggaa gagaaagaag cagaaggtta ctttggcgag cttattaaca gaggatggat 6000
tacgcagcac ggagacaaca acagttataa ttactatgag atccaccccg tgatgctggc 6060
cttcctgaga tgcaagtcca aggagtacaa ttttttaaca tgcttgggtc tgggatctga 6120
tactagtact agtgcatcct ccccaaggtt gattcgccgg ctgtctcttc agggggggta 6180
tccagtggac tgcttgtcaa gcatgagtat ggatgtgtca cacacttgca gccttgtcgt 6240
ccttggtgac gtggcgcgac ccaagggaat ccccttctat atgtttaagc gcttgcgagt 6300
gttggacctt gaagataata aggatataca ggattctcat ctgcagggca tatgtgaaca 6360
gttaagcctc agagtgaggt accttggtct caagggaacg cggatccgaa agctccctca 6420
ggagatgagg aagctgaagc atttggagat tttgtatgtg gggagcactc ggatcagtga 6480
acttccgcaa gagattggag agctgaagca tctgcggatt ctggacgtga gaaacacgga 6540
catcactgag ctcccactgc agatacggga gctgcagcat ctgcacactc tggacgtgag 6600
gaacactcca atcagtgagc tcccgccgca ggttggcaag ctgcagaatc tcaagattat 6660
gtgcgtgagg agcactgggg ttagggagct cccaaaggag attggggagc tgaatcatct 6720
acagactctg gacgtgagaa acacgagggt gagagagctg ccatggcaag ctggccagat 6780
ctcccaatcg ttgcgcgtgc ttgccggtga cagtggcgat ggcgtgcggt tgcccgaagg 6840
cgtctgcgaa gctctgatca acggtattcc aggggctacg cgtgcaaaat gcagggaggt 6900
tctgtccatc gcgatcatcg atcgtttcgg acctcccctt gttgggatat tcaaagttcc 6960
cggcagtcat atgcgtatcc cgaagatgat caaagaccac ttccgcgttc tttcttgcct 7020
agacatcagg ctctgccaca agcttgagga tgatgaccaa aagttcctcg ccgagatgcc 7080
caacctgcag acgctcgtgc tgaggttcga ggccctacca agacaaccca taaccatcaa 7140
cggcacaggc ttccagatgc tggagagctt ccgtgtcgac agccgggtgc caaggatagc 7200
cttccatgaa gacgccatgc ccaacctcaa gcttctcgag ttcaagttct acgccggccc 7260
agcaagcaac gatgccatcg gcatcaccaa cctgaagagc ctccaaaagg tggtctttcg 7320
gtgctcgcca tggtacaaga gcgacgcccc tggcatcagc gccaccattg acgtcgtgaa 7380
gaaagaagcc gaggagcatc ccaaccggcc gatcaccctc ctcatcaatg ctgggtataa 7440
ggagatatca actgagtcac acgggagcag tgaaaacatt gcgggcagca gtgggatcga 7500
tactgagcct gcacaggcac agcatgataa tctccctgct gttcgagatg actacaaggg 7560
aaaagggatt cttcttgatg gcaggtgtcc tacctgcggc cgagcgacta aaattgaaga 7620
ggaaacccaa gatcgagtag cagatattga aattcaaaca gaaactacta gctagctagt 7680
accgctgtct aattttattt gtattaaata ctctccttca tgtaatactc taagtccccg 7740
ctcaattttt cttgcctccg caactctaca tgatactcaa aactgatttc ttacctccct 7800
ttgattgtag aaaaaacatg ggaagtttgg aggatttcaa tcctatgaaa aggactttga 7860
gaaacaaatt acggaatcct atcttttcct ttaaaagata tataatagaa aatcctatag 7920
agatttttat aggaaagtta acaaaagcct caatcttatg gagaattcct gtgtttattt 7980
gcatagagga aagtgagact cacctaatca ggggcggttc tagtgggtgg tccaggtgat 8040
ccctggacca ccctaaaatt tggcccatga gaaccgcccc ccccctcctc ctcccggggt 8100
tggtcgaaac cagaaaaaag gccactgagc taagctaggg ttagggagag gtgtagccat 8160
gaactagcca cagcagcacg catcgctggc tgccgcctcc gcctctgcgt cagcgccagc 8220
gtgccgtctc cccggctctc gactctgcct ccgcctccgc cttggcgccg atgcgtcgcc 8280
tcctcggctc ccgcttctac ctcagcgcca gcgtgccgcc tccgccttag cgccgcctcc 8340
tcgcctcctc ccttccccga ctcctccagt cccccactcc tgtcggttgc tatcgtcggc 8400
gggcggcgac caacgcctac aagccaacta cggctgcctg ggcaagccgg cggccacccc 8460
cgcccctgga aagggaaaac attgccattc cacaggcggc gccaggtccc ctttctctct 8520
agctttcttg ttgctcagca aagttcaaag gtcagtagac atttcttgtt ggttttttgc 8580
aatgttgccc tatgcagtag tgagtagtca gtactcagta gtagtgctag tgaaaaattt 8640
ctgccttgag tgtagtgaaa agcaagctga gaaatagcat gtgtgacaaa ttattaaatg 8700
atggtttaat cacatctatt gagcgggtta ttttctctca agttagcaaa gaagacatta 8760
tgcataattt catgtctata tatgaacgga gagtagaaaa gaagtagggt tttgtaatct 8820
tctacttatc tattataaga tatttgaatg attatgttct aactttcgta acttcaaatt 8880
tagatgttgg ttttaagcaa gttgtgatgt tgtgagatat attgctacat ttcaattata 8940
tttttattat ttttgacaca tttggcttga atttgtaaca atgtgacaat ttggagcatc 9000
aagtaggact acccgacgat tcatgaaacc aatgtgccga tatttagcgg ttcttaatca 9060
tactgaggta agaaggcaac attttgcaaa agtagaattt cttcatgaaa atcaggctta 9120
tagctttaat gttcacaaac tagctaagtt tccttgtacc ttagaggagg ggcatcatgt 9180
tcgattctca aatactcctt gtaatctaag taggaatgta aataatttgg ttgttaatga 9240
ataaaatgcc agaattttat tctaaaagaa tatgtaattc aaattaatat tattctagat 9300
gtatcttgaa tagcccatta gcactggacc agtctttcaa gttgggtcag tggtcggtac 9360
cattgcctgg cccatcatgg aacagaatgg ccatggtctg atcgcaatgg gccgaggcat 9420
gacgcgctgc gctttcgaaa tcacgggcct gagctgtcat gagggctatc ctgggccagg 9480
cccgcgagac aagagggtgt cggacaatgc aacaacaacg agaggcaata agcagagctt 9540
atgcattgca cgaccaacaa agttgtcact tacatccctt attattctca atcttaatac 9600
tatctccgtt caaaaaaaag tagactttta gatacgaatg tgaacaaacg tcctcatcaa 9660
tcttgcctgc tgataaagca accgcagcta ccacgcacta tgcagggatc gatcgatcga 9720
tggatggaca tattattact caacaaagta gaggcattaa tccgcttgca gactaatcag 9780
attcaggcga cagcacgcgg ctgctgattc atcagatggc attgcttcaa attgttcgct 9840
ttcaccatgc cctgcaccat gcatcattta gatatttatt ttactgtact catagtaatt 9900
aaccactgaa aattcactct aggaaatcgt gtgttaactg ttaacctggt agattttgat 9960
catgaattgg ttgatgtcgt cgagcgatat gacgccctcg tacgcggcgt tgaaacggta 10020
gctgtcgtca tcgcgtccag acgccaccag cagcggcggc gcaggcggca gctgctccac 10080
gcagctgtac tcgcactcgt cgtggcactc gccgtcgccg ccgccgccgc cgccgccgtt 10140
ctccgccgtg atcgccacct gcagcgtcag gtgcacctcc agcgtcacga acacgcccac 10200
caccttcgcc ggcgacccca tcgccgtcgg ccgctgcatc ttgtagctcg acatccacga 10260
gaatatgtac gccgccatcg acgacgagac gccgacgatg ttctgcttcc tcaggctgtt 10320
c 10321
<210> 6
<211> 8839
<212> DNA
< 213>Oryza paddy rice (Orysa sativa L.)
<220>
<221> promoter
<222> (1)..(1209)
<220>
<221> 5'UTR
<222> (1210)..(2441)
<220>
<221> CDS
<222> (2442)..(3433), (3598)..(5671)
<220>
<221> 3'UTR
<222> (5672)..(5915)
<400> 6
ctactctctc acctcctgac tctccgactc cgagtgtaat tagagcaagg ttaacagtac 60
agccaagtag tagcttcaat ttatttatag ccaatctaat agcttattca tacaagtact 120
atactattaa tacctggtcc cacctgtcat acacactgtg tcttggagtc tgtgctacag 180
ttggctacaa atctatagcc cgctaccctt ctctctcctt atttatctcc ttaaaatatg 240
tttgcagttg gcttatagcc tgctattgta actgctctta aagtggtggt atttttctcg 300
gtcctctcgc aatgggttag agaactgctt tcacataatt agttcgtttg ttctctaata 360
taaggtattt taacatttta ctttgtactc ccttcattct aaattgatct atatatagtt 420
tttttaggtt attcctaaat gatctatata tttgtgttca tttattaagt ctattcgtta 480
tttgtgtgca ttggagtaaa tggacattga tgcatgcttc catgcacaca agtatttata 540
acccacatgc aatatcttga tttgctattg gctaggaaat attggggatg atgcatgcat 600
tgagtttgtt gttagagtaa atataatatg aaagagttat tagtttttct tggtcttggt 660
gtacctatga aatacgtaga tcaatttaga atggagggag tacgatcctt cgttttatta 720
aaaaaaagtt tgaaaattat tatttaattt tcttttactt attttattat ctaaaatctt 780
ttaagcacaa ctttttattt ttattatttg cataattttt ttaaaataag ataaatggtt 840
aaataggact ccctctattc cataatataa ggcacaacca cttttcttag atgtttcata 900
atataaggca tgcatgcata taggcaatta actatgacct cttttttatt aaattattat 960
tcatgctctc tgattctatt ggatgcatgc attgtattta ttaggatgtt ctaaactaca 1020
agataataat aattattttc ttggtgtttg ggttaggggt ggttatgact tatattttat 1080
aatggaggga gtagtacaag tataatatta aagactttaa tattatatta aaggacggag 1140
gaaatattag ggagaaacgg catgaccacg agtcagtcaa cgagtctgcg aatgcagtgc 1200
agtattattg gcctccgccc aaaagcctca cccccaaggc cccaatcttc ccaccgcgcc 1260
gctgtccccc cccccccccc cccccccaat cttctggcct ggccagctct ggtcggagga 1320
gggttcagtt ccatgggtgc gccccctgcc catccctcca cttctacttt cgtcttgaga 1380
tctcgccgtt tctgatctcc ttgccatggc aaccagttga tttggctgcc aagatcgtcc 1440
gtgctttgca aaggtgaaca atctttttct tcttgggctt tcactttgtt gtgttaatag 1500
actcagcgcc gtcgcaactt gcaaataaaa aggttcagat cggaaaatgg aaataccgta 1560
cacagtagtc tatagagaat cgactagtat ttcctgactg taagtaagga agttgtaggt 1620
accatgttct gatgattatc tttgaagctt ggagggggac aaaaagttgg gagagaaaca 1680
agaatgtatt tgttcacttg attgggggca gtaggtaaaa tctcagcgtt aggaatttgc 1740
acctctttat taagtttgac taaatttata gaaaaaaatt agcaacactt aaaacaccaa 1800
attagtttca ttgaatccaa cattgaatat attttgataa tatgtttact ttgtgttcaa 1860
aatgttacta tatttttcta taaatttgat caaatttcaa taagtttgac tagaaaaaaa 1920
agtcaaaccg acttataata tgaatataaa tggagggagt agattataaa ttatctttat 1980
tataatactt tgagtgtaca aacgaatgaa taaagtttac aaagttgaaa agttattttt 2040
aaatactatt ccgataaacc tgtatataga gaacttgtga aaaaaaacac cgtttgaaat 2100
gtgtctgtga taatctataa gaggaacggg gcctaaatat attccttttt tcctacgaaa 2160
atgcaagaag tattgcctaa tatattgata gagctgaagt tttaaagtaa aatgaaacat 2220
ttgcatgagg cagcactgga caaaaagcgc ttgtgaaaaa aatatgttcc aaatattatt 2280
aattcattca tggtatggtc tttattttca ttttcccccc ttatccatgg gcaggacacg 2340
gaaattctca gcaggttcgc gcttgatctg aactgtctgt ggctcatact ctcttgtgct 2400
tgcaacagct gaaagttgca gtgagaagta cagagaacaa gatggagttg gtggtaggtg 2460
cttccgaagc caccatgaaa tctctcttgg gcaagctggg caatcttcta gcccaggagt 2520
atgctctcat cagcggtatc cgtggtgaca tccagtacat caatgacgag cttgccagca 2580
tgcaggcctt cctccgtgat ctcagcaacg tgccagaggg tcacagtcat ggccaccgga 2640
tgaaggactg gatgaagcag atccgagaca tcgcctatga tgttgaggac tgtatcgatg 2700
actttgccca ccgcctccct caggattcca tcagcgatgc caaatggtcc ttcctactca 2760
caaaaatcta tgaactatgg acatggtggc cacgtcgtgt gattgcttcc aacattgccc 2820
aactcaaggt acgggcacaa cagatcgcag atcgacgtag tagatacgga gtgaacaacc 2880
cagaacacct tgacagtagc agcagtgcca ggacccgtgc tgtcaattac gaaattgctg 2940
agtatcaggt cacaagccct cagatcattg gtataaagga gcctgtgggg atgaagacgg 3000
tcatggagga gcttgaggtt tggttaacta atcctcaagc tgaaaatggg caagctgttc 3060
tgtccatagt cggttttgga ggtgtgggaa agactaccat tgccacagca ttgtacagaa 3120
aagtcagtga taaatttcag tgccgggcat cagtagctgt gtctcagaac tatgaccaag 3180
gcaaagtcct caatagtatt ctgagtcaag tcagcaatca ggagcagggc agcagcacaa 3240
caattagtga gaaaaagaac ctcacctcag gcgctaagag catgttgaag acagccctgt 3300
cactgctcag aggtaattgt atatgtcagc cagaaaatga tggaaaccct gataatacac 3360
caatcaggct gcaggaaaca acggacgatg atcaaaaccc cagaaaactg gaacagctcc 3420
tggccgaaaa gaggtacctt tttttgtaaa taaaattgct ttgcttatct gtaaattaac 3480
ttactcatcc cactctaaat ctaatgttta tttccttcta tacacagcac aactccatct 3540
tttgaatggg ttttattttt ctcacttgtg ctcatttttt ttttatcatc tctgcagtta 3600
tatcctcttg attgatgaca tttggtctgc cgaaacatgg gagagtatca gatcgatttt 3660
gcctaaaaat aataaaggcg gtagaataat agtgactaca agatttcaag ctgttggttc 3720
aacatgctcc cctcttgaaa ctgatcgttt gcatacagtt gattttctca ccgatgacga 3780
gtcccaaaac ttattcaata caagtatttg tgaatcaaag ataagaaaag atagcaacaa 3840
agtagacgag caagtccctg aggaaatatg gaaaatatgt gggggattgc ctttggccat 3900
agtcaccatg gctggtcttg tcgcctgcaa cccaaggaaa gcctgctgcg attggagtaa 3960
actttgcaaa tcattatttc cagagcaaga aactcctctt accctcgatg gtgttacaag 4020
gatactggat tgttgttaca atgatttgcc tgcggatctg aagacttgct tattgtactt 4080
gagtatattt ccgaagggtt ggaaaattag taggaaacgt ttgtcccggc gatggatagc 4140
tgaaggtttt gctaatgaga agcaagggtt aacccaggaa agagttgcag aggcatactt 4200
taatcaactc acaagaagga acttagtacg tcccatggag catggcagca atgggaaggt 4260
aaaaacgttt caagttcatg acatggttct tgaatacatc atgtccaaat caatcgaaga 4320
gaattttatt actgtggttg gtggacactg gcagatgact gcaccaagca ataaagtccg 4380
tcgactgtcg atgcaaagca gtggatccaa tcgtggaagt tcaacaaaag gcctgaactt 4440
ggctcaagtg agatcactga cggtgtttgg gaacctgaac catatgccat tccattcatt 4500
caactatggg ataatacagg tgctggatct tgaggactgg aagggtttga aagagagaca 4560
tatgacggag atatgtcaaa tgcttttact caagtatttg agcatccgac gaacagaaat 4620
ttccaaaatt ccctccaaga ttcagaaact tgagtacttg gaaactcttg acataaggga 4680
gacatatgtc agggacctgc ctaagtcaat agtccagcta aaacggatca ttagcatact 4740
tggagggaat aaaaacacac ggaaggggct gaggttgcct caagaaaaaa gtaagaagcc 4800
aattaaaaac ccgtcgcctc aaggaaaaac aaaggagccc gcaaagaaag gattcttatc 4860
ccaagaaaaa ggtaaaggcg caatgaaagc actccgtgta ctgtcaggga ttgagattgt 4920
tgaggaatca tcagaagtag ctgcaggcct tcatcagttg acagggctaa ggaagcttgc 4980
catatacaag ctcaatataa caaagggtgg tgataccttc aaacaattac agtcctccat 5040
tgagtacctt ggcagctgtg gtctgcagac tctggccatc aatgatgaga attctgaatt 5100
tatcaactca ctgggcgaca tgcccgcgcc tccaagatat cttgtcgccc ttgagctgtc 5160
tggcaagttg gagaagctac ccaagtggat caccagcatc actactctca acaagctaac 5220
catatctgta acagttctta ggactgaaac tttggagatc ctccacattt taccttcatt 5280
gttttccctc accttcgcct tttcacttag tgcagcgaag caggatcagg acataataaa 5340
ggacatcctt gagaataata aattggacag tgatggggaa atcgtcattc cagctgaagg 5400
attcaagagt cttaagctgc ttcgcttctt tgcaccttta gtgccgaagc tcagcttttt 5460
ggacaagaat gcaatgccag cactcgaaat cattgaaatg cggtttaaag acttcgaagg 5520
tctatttggc atcgaaatcc ttgaaaatct ccgtgaggtg catctcaaag ttagtgatgg 5580
ggcagaagca ataaccaagt tccttgtaaa tgatttgaag gttaatactg agaaaccaaa 5640
agtatttgtt gatggcatcg tcactgcatg agaagtaaaa ttgctgcaaa tcggagaact 5700
taccaatcat ctgaggcttc ccctctatta ttactctctt agaatatatt gttattattg 5760
ctcaccttgc aaaataaaat agggatggca tagcatattg ctacaacgta ccatggttcc 5820
atcatagttg atttcacttg tcattacagt gtctgttcag ttgtgttttc tattaataaa 5880
agggagatct ccgcaagaaa ccattattat acttatattc ggttattgaa ctctataaat 5940
gatgggattg ctatatttat ggtgccacaa tttccatgag tgcggtattt ttttttctag 6000
cagttggctg gtgttaagat ttgtgctgcc attgctcctc tattattggt gcccaaaatt 6060
acgctttgca ctatgttcac agttgtaaaa cttacctaaa tttcgtgtat tagtactgta 6120
atgttgtgat tttcgcgtcc agttatattt ttttctttgc cagaagtttg atttcaaggt 6180
atatctggta agcttcagcc ggattcgtta agtttttagt ttaatcagct aaacttttaa 6240
cagtatgcat gcactagttt ctcccgttat cttatccaat ataattaatg ggctaattgg 6300
atctatgcca ttacaaacat ggcaagattc agaaaaatgc tactacaatt tgtctattca 6360
tagctgtgct agtgaaattt tacaaaattg gaaccgtgct attgatatca cgttttccat 6420
ccatcttttc ttttttttcc ttttcttctt tatcccgtct tcttcccgca ccgacgagag 6480
gcgagtggcg gcgggaccgg caagcgatgg caggacctcg gcagctcagt ggtggcagcg 6540
gcaaaaggga aaggggaagg ggtaggagag aacctggccc gccgacgcaa gcatgttggt 6600
cgtcgtcttc cccgaggagc ttgagcagtg gcaatggtag aggaatccaa ggaggccgag 6660
gaggttgagc ggcactaatg gtagaggatg ccgaggaggt cgagcatcag cgagcttgcc 6720
ctgggtgggc gagtgaggga gagctctccc aagtgtctgc gaagctctca tcttcgtttt 6780
cctcatcccc actttgctcc tcttccacta ctgctgttga agctagcggc cctctctcac 6840
cagatctggg cggagctcaa gcgaggcgcc accaccgacg atggagcttc gcgtggaccc 6900
cacagcctca cccgattgcc atccctccta ccctcatcac tccatctcca agcccaaatc 6960
ccacctcccc caccggctcc tcctcctcag cccctgccgt ggcagccgag gccaccgatg 7020
ggccaaccct caacatcgtc tcgaagcagc tccgcgcact gtggaagaag cacaaccaaa 7080
tcctccagat ggaggagtcg ctcactggcg ggaggaagct gaacaaggag caggaggagg 7140
tgctccgatc caagcccatc gtcatcgcgc tcattgatga gctcgagcgg atgcgtgccc 7200
tgctcgccga cgccctcgcc gaggagctct cctcctgccc cgccccttcc ctatctgtca 7260
ctccctcctc ctccacctcc tccggcgctg atttgtccgt caaggatctc ctcacgctca 7320
tctacttcgg ctccctcttc gacgtcaagt cgcagatcga gttcgtcacc accatgctcg 7380
cacgctaagg agctagactg atgcatcacc tacgactatg tccgtccttg ccgcagcacg 7440
acttgctacg gtcccgcctc acgccccgtc gctcgccggt cccgccgcca cttgcttctc 7500
atcggtgcgg gaagaagatg ggagaaagaa ggaagaaaac gggaaaaaaa gagatggatg 7560
aaaaacgtga cagcaatagc acggttccaa tttcgtaaaa tttcaatggc accgctacga 7620
atagaagaat tgtagtagca tttttctgaa tcaacatatt tgtaatggca tggatccaat 7680
aaaccccaaa tttattttat gccaatacaa atattggcaa ctggtgtcat taacgtataa 7740
atgggtcgcc tacataaaac cttccgcgtc cttcttggtt gaacaggtaa ttaattgcaa 7800
ccattttctg aaaaaattga ctggaagtct gaaaccgtcc ctgctactcc ctgaatgctt 7860
tagttcaatc tctatgaagc tccatccaaa caaaatctcc agatgatatt tcattaaaaa 7920
catcatgtcc agaactgtcc agtatccaag aatggctatg atcttttctg aattctggac 7980
tcgggactta tattacatgt atcaaatcca gttattttct ggagctacac cagatgaaat 8040
caccaagtaa aacaattata taaggatggt gagttgttaa ttaattacag ctttgataga 8100
ccactctgtt tgttaatgcc tgacaagagt tcacaaagaa aaaaatacac aggattacaa 8160
aactttttag atactatctg atgtgtgtta attctctcaa atactatgta cctgaacaaa 8220
caagagagaa catgttgtga attgtgatct cctcatgcga gtgacctgaa actctctaca 8280
gggtcagact cctcaagcat cactgcagcc tggaaacaat cagcagcatc attgatcctc 8340
ccatcattcc tgtgaactct ccctaaatga agccaagcca tcctgttcgt aggctcaatt 8400
ctcagtgcat cagagaggaa gcatctcgct gcagggagat accttgaccc ttgcttgcag 8460
agaagtgcac caatggccac cttggatgga acatgctcta gctctatcga gaatgcgttg 8520
acgtaggcag ccagtgcctc cttgttctga tctcgcgcct cgagcatgaa acctgaactt 8580
atagcatcca cgagtcagat tttgatgttg aattcagttg aatgcagaga attgtcatgt 8640
tagtgttaca tagaagtggt gaaagttcaa gtttagttga gttgatcacc ttctgcatgc 8700
attgttgcag ccgagtagga ttttagggct ctagccttct gcagacatat ctcagcgtct 8760
ctccagattg agagactgga gtacaggttt gcaagtcctt gccatatttc aaactcactt 8820
acactgtcat tctgtccct 8839
<210> 7
<211> 38
<212> DNA
< 213>artificial sequence
<400> 7
ttttggcgcg ccggtgcggg aagaagacgg gataaaga 38
<210> 8
<211> 38
<212> DNA
< 213>artificial sequence
<400> 8
ttttggcgcg ccgccgacga tagcaaccga caggagtg 38
<210> 9
<211> 23
<212> DNA
< 213>artificial sequence
<400> 9
gaagctctga tcaacggtat tcc 23
<210> 10
<211> 23
<212> DNA
< 213>artificial sequence
<400> 10
tctttgatca tcttcgggat acg 23
<210> 11
<211> 23
<212> DNA
< 213>artificial sequence
<400> 11
tgaacttcca cgattggatc cac 23
<210> 12
<211> 25
<212> DNA
< 213>artificial sequence
<400> 12
acatggttct tgaatacatc atgtc 25
<210> 13
<211> 18
<212> DNA
< 213>artificial sequence
<400> 13
gcaggtcagc caagcaat 18
<210> 14
<211> 19
<212> DNA
< 213>artificial sequence
<400> 14
caaccgttgt tttgcctcc 19
<210> 15
<211> 20
<212> DNA
< 213>artificial sequence
<400> 15
<210> 16
<211> 20
<212> DNA
< 213>artificial sequence
<400> 16
<210> 17
<211> 12
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 17
Gly Leu Pro Gly Gly Gly Lys Thr Thr Val Ala Arg
1 5 10
<210> 18
<211> 12
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 18
Asn Lys Lys Tyr Leu Ile Val Ile Asp Asp Ile Trp
1 5 10
<210> 19
<211> 15
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 19
Asp Leu Gly Gly Arg Ile Ile Met Thr Thr Arg Leu Asn Ser Ile
1 5 10 15
<210> 20
<211> 20
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 20
Glu Asp Ser Pro Cyc Tyr Asp Ile Val Asn Met Cys Tyr Gly Met Pro Leu Ala Leu
Ile
1 5 10 15
<210> 21
<211> 16
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 21
Val Leu Ser Ile Val Gly Phe Gly Gly Val Gly Lys Thr Thr Ile Ala
1 5 10 15
<210> 22
<211> 18
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 22
Leu Glu Gln Leu Leu Ala Glu Lys Ser Tyr Ile Leu Leu Ile Asp Asp Ile Trp
1 5 10 15
<210> 23
<211> 13
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 23
Gly Gly Arg Ile Ile Val Thr Thr Arg Phe Gln Ala Val
1 5 10
<210> 24
<211> 20
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 24
Glu Gln Val Pro Glu Glu Ile Trp Lys Ile Cys Gly Gly Leu Pro Leu Ala Ile Val
1 5 10 15
<210> 25
<211> 13
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 25
Cys Leu Leu Tyr Leu Ser Ile Phe Pro Lys Gly Trp Lys
1 5 10
<210> 26
<211> 13
<212> PRT
< 213>Oryza paddy rice (Orysa sativa L.)
<400> 26
Lys Thr Phe Gln Val His Asp Met Val Leu Glu Tyr Ile
1 5 10
Claims (8)
1. rice blast resistance gene
Pi7, its nucleotide sequence is shown in SEQ ID NO:1.
2. rice blast resistance gene according to claim 1
Pi7, it is characterized in that its amino acid sequence coded is shown in SEQ ID NO:3.
3. claim 1 or 2 said rice blast resistance genes
Pi7The cDNA sequence.
4. claim 1 or 2 said rice blast resistance genes
Pi7Encoded protein matter.
5. by claim 1 or 2 said rice blast resistance genes
Pi7Or the expression cassette of said cDNA sequence of claim 3 and promotor structure.
6. contain claim 1 or 2 said rice blast resistance genes
Pi7Or the expression vector of the said cDNA sequence of claim 3.
7. claim 1 or 2 said rice blast resistance genes
Pi7, the said cDNA sequence of claim 3, the said protein of claim 4, the said expression cassette of claim 5 or the application of the said expression vector of claim 6 in the preparation transgenic plant.
8. claim 1 or 2 said rice blast resistance genes
Pi7, the said cDNA sequence of claim 3, the said protein of claim 4, the said expression cassette of claim 5 or the application of the said expression vector of claim 6 in improving the rice anti-rice blast resistance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105786332A CN102094027B (en) | 2010-12-08 | 2010-12-08 | Rice blast resistance gene Pi7 and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105786332A CN102094027B (en) | 2010-12-08 | 2010-12-08 | Rice blast resistance gene Pi7 and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102094027A CN102094027A (en) | 2011-06-15 |
| CN102094027B true CN102094027B (en) | 2012-11-14 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010105786332A Expired - Fee Related CN102094027B (en) | 2010-12-08 | 2010-12-08 | Rice blast resistance gene Pi7 and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102094027B (en) |
Families Citing this family (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102732511A (en) * | 2012-03-08 | 2012-10-17 | 华南农业大学 | Functional specificity molecule marker Pi7FNP of blast-resistance gene Pi7, method and application thereof |
| CN102732531B (en) * | 2012-03-23 | 2013-10-09 | 南京大学 | Rice blast resistant gene RMg7, RMg8 or RMg9, and its application |
| CN102747090B (en) * | 2012-07-02 | 2014-06-11 | 中山大学 | Rice disease resistance gene OsLYP4 and application thereof |
| CN103409433A (en) * | 2013-08-12 | 2013-11-27 | 江苏省农业科学院 | Novel broad-spectrum rice blast resistance allele pi21t and resistance application thereof |
| CN103421841B (en) * | 2013-08-15 | 2015-04-15 | 中国农业大学 | Os74 protein for rice and application of Os74 protein |
| CN104404052B (en) * | 2014-09-11 | 2017-01-25 | 南京大学 | Rice blast resistance gene RMg39 and its application |
| CN104293799B (en) * | 2014-09-11 | 2017-05-17 | 南京大学 | Rice blast resistance gene RMg38 and application thereof |
| CN104372011B (en) * | 2014-09-11 | 2017-01-11 | 南京大学 | Rice blast resistance gene RMg41 and applications thereof |
| CN104845977B (en) * | 2014-12-22 | 2017-04-19 | 广东省农业科学院植物保护研究所 | Rice blast resistant gene Pi50, preparation method and application thereof |
| CN106480051B (en) * | 2015-08-24 | 2020-06-19 | 中国种子集团有限公司 | Recombinant nucleic acid fragment RecCR010113 and detection method thereof |
| CN109280082B (en) * | 2018-10-19 | 2019-06-11 | 云南省农业科学院农业环境资源研究所 | A kind of rice blast resistance gene Pi57 and its coding albumen and application |
| CN113122649A (en) * | 2021-04-09 | 2021-07-16 | 武汉科技大学 | Primer and method for screening new allele of rice blast resistance Pik locus |
| CN113744800B (en) * | 2021-06-09 | 2022-06-24 | 华南农业大学 | Technical system with inclusion and accurate identification and excavation of rice blast Pik disease-resistant allele family, application and molecular marker |
| CN113789403B (en) * | 2021-09-02 | 2022-09-06 | 华南农业大学 | Three sets of compatible and accurate rice blast Pid disease-resistant gene family allele identifying and mining technical systems |
| CN116640774B (en) * | 2023-06-14 | 2024-02-09 | 江苏省农业科学院 | Rice blast resistance gene Pik-W25 and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1844393A (en) * | 2006-03-06 | 2006-10-11 | 华南农业大学 | Resistance gene Pi37 against rice blast and use thereof |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1844393A (en) * | 2006-03-06 | 2006-10-11 | 华南农业大学 | Resistance gene Pi37 against rice blast and use thereof |
Non-Patent Citations (4)
| Title |
|---|
| Campbell MA et al.Debelopment of co-dominant amplified polymorphic sequence markers in rice that flank the magnaporthe grisea resistance gene Pi7(t) in recombinant inbred line 29.《Phytopathology》.2004,第94卷(第3期),302-307. |
| Debelopment of co-dominant amplified polymorphic sequence markers in rice that flank the magnaporthe grisea resistance gene Pi7(t) in recombinant inbred line 29;Campbell MA et al;《Phytopathology》;20040331;第94卷(第3期);302-307 * |
| 冯淑杰 等.稻瘟病菌新无毒基因AvrPi7的遗传及物理作图.《科学通报》.2007,第52卷(第3期),283-290. |
| 稻瘟病菌新无毒基因AvrPi7的遗传及物理作图;冯淑杰 等;《科学通报》;20070228;第52卷(第3期);283-290 * |
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