CN101792748B - Paddy istone lysine methyltransferase, coding genes thereof and application thereof - Google Patents

Abstract

The invention belongs to the technical field of biological histone genes, and particularly discloses paddy histone lysine methyltransferase, coding genes thereof and application thereof. The methyltransferase has one of amino acid residue sequences, namely 1) an amino acid residue sequence SEQ ID No:1 in a sequence table; and 2) an amino acid residue sequence formed by substituting and/or losing and/or adding 1 to 50 amino acid residues in the amino acid residue sequence SEQ ID No:1 in the sequence table. The methyltransferase is a protein that has the regulation and control effect on the growing development of plants. Antisense transgenic strains of the coding genes of the histone lysine methyltransferase of the paddy are short and small in growth. The histone lysine methyltransferase provides the high-quality genes for the improvement of plant varieties, and has high practical application value and wide application prospects.

Description

The histone-lysine methyltransferase of paddy rice and encoding gene thereof and application

Technical field

The invention belongs to biological histone gene technical field, be specifically related to a kind of histone-lysine methyltransferase relevant with the paddy rice epigenetic regulation and encoding gene and application, particularly this albumen and gene thereof the application in regulating growth of plants.

Background technology

All the transcriptional expression regulation and control with gene are relevant for grow and the process such as stress response of plant.And the regulation and control of the transcriptional expression of gene are except being subjected to inherited genetic factors to affect the regulation and control that also are subject to epigenetics (epigenetics).In recent years, epigenetics became the hot fields of life science already.Except dna sequence dna, also have the information of many regulatory genes in genome, they itself do not change the sequence of gene, but can pass through genetic modification, thus function and the characteristic of impact and regulatory gene, and by cell fission with breed heredity and go down.Histone, as the chromatin fundamental unit--the core component of nucleosome, its modification are that important epigenetics one of is modified.The Histone Code (histone code) that study group is protein modified and these various combinations between modifying form will be the long-range mission of epigenetics research to the regulatory mechanism of all great processes of vital movement.

Methylating of 36 Methionins of histone H 3 N-terminal (H3K36) is considered to activate relevant with genetic transcription usually.Yet its research to the regulating and controlling effect of growth and development of plants just just began in nearly 2 years.With only have one or several histone H 3 K36 methyltransgerase (histone lysine methyltransferase, HMTase) difference in fungi and the animal, generally have a plurality of H3K36 methyltransgerases in the plant.Note according to chromDB (http://www.chromdb.org/), the protein of 5 genes encodings and the H3K36 methyltransgerase homology in fungi and the animal are arranged in the arabidopsis gene group, that study at present at most is SDG8 (Zhao Z, Yu Y, Meyer D, Wu C, and Shen W-H.Prevention of early flowering by expression of FLOWERING LOCUS C requires methylation of histone H3K36.Nat Cell Biol, 2005,7,1156-60.).SDG8 specifically the two of catalysis H3K36 methylates and trimethylammonium, the sdg8 mutant is because the disappearance of SDG8, the H3K36 methylation level that causes FLOWERING LOCUS C (FLC) to comprise the chromatin zone of important promotor of transcribing element and First Intron descends, and finally causes the plant early blossoming thereby cause the FLC expression level to descend.Arabidopis thaliana SDG8 is also inferred to have different physiological roles except affecting flowering time.The sdg8 mutant also has multiple phenotype except showing the early blossoming phenotype, comprises that plant obviously diminishes, degradation under the fertility.The analysis of Microarray further show SDG8 may the multiple physiological pathway such as transcriptional regulatory, signal transduction, internal metabolism have potential function ( Xu L, Zhao Z, Dong A, Soubigou-Taconnat L, Renou JP.Steinmetz A, and Shen WH.Di-and tri-but not monomethylation on histone H3lysine 36marks active transcription of genes involved in flowering time regulation and other processes in Arabidopsis thaliana.Mol Cell Biol, 2008,28,1348-60.).

Arabidopis thaliana SDG8 affects on what grow whether their homologue has the function of no less important in the let us thinking paddy rice.Different from the dicotyledons Arabidopis thaliana, paddy rice is a monocotyledonous Typical Representative.Dicotyledonous and monocotyledons, although may have higher conservative property between gene, may there be very big-difference in their regulatory mechanism that grows.Owing to reasons such as the paddy rice life cycle are long, correlative study also quite lags behind with respect to Arabidopis thaliana, the histone H 3 K36 at present blank out especially of the research of modifying that methylates in the paddy rice.And the value of Study On Rice not only is the economic worth that it is huge, is that also it is the ideal model that research is all other crops such as unifacial leaf grass corn, wheat, Chinese sorghum.

Summary of the invention

The purpose of this invention is to provide histone-lysine methyltransferase and encoding gene and an application that derives from paddy rice.

Histone-lysine methyltransferase provided by the present invention, name is called SDG725, derives from paddy rice (Oryza sativa ssp.japonica), is one of following amino acid residue sequences:

1) the SEQ ID No:2 in the sequence table;

2) with the amino acid residue sequence of the SEQ ID No:2 in the sequence table through replacement and/or disappearance and/or the interpolation of one to 50 amino-acid residue and growing of plant had the protein of regulating and controlling effect.

SEQ ID No:2 in the sequence table is comprised of 2150 amino-acid residues.

The gene (SDG725) of code book invention histone-lysine methyltransferase is one of following nucleotide sequence:

1) nucleotide sequence of the SEQ IDNo:1 in the sequence table;

2) DNA of the SEQ IDNo:2 protein sequence in the code sequence tabulation;

3) with sequence table in the nucleotide sequence that limits of SEQ ID No:1 have 90% above homology and the nucleotide sequence of the identical function protein of encoding;

4) under the rigorous condition of height can with sequence table in the nucleotide sequence of the dna sequence dna hybridization that limits of SEQ IDNo:1.

The rigorous condition of above-mentioned height is: (or the solution of 0.1 * SSC), 0.1%SDS is hybridized under 65 ℃ and is washed film with 0.1 * SSPE.

SEQ ID No:1 in the sequence table is by 6453 based compositions, and its encoder block is that coding has the protein of the amino acid residue sequence of the SEQ ID No:2 in the sequence table from 5 ' end 1-6450 bit base.

The primer pair that contains arbitrary fragment in recombinant expression vector, transgenic cell line and the engineering bacteria of gene of the present invention and this gene that increases all belongs to protection scope of the present invention.

The present invention also provides the application in regulating growth of plants of above-mentioned histone and encoding gene thereof.

In actual applications, with the antisense expression vector rice transformation of above-mentioned paddy rice histone-lysine methyltransferase gene, obtain the short and small paddy rice of plant strain growth, growing of plant is regulated.

Above-mentioned recombinant expression vector all can make up according to ordinary method.

The antisense transgene plant strain growth of paddy rice histone-lysine methyltransferase gene of the present invention is short and small.The present invention provides a Fineness gene for the improvement of plant variety, simultaneously also to deep biological function and the biological procedures of understanding histone-lysine methyltransferase, further decode Histone Code and understand that Histone Code is grown higher plant, the effect in the heritable variation has very important significance.Protein of the present invention and encoding gene thereof have higher actual application value, have a extensive future.

Description of drawings

Fig. 1 is the protein-active measurement result of paddy rice histone-lysine methyltransferase gene SDG725.

Fig. 2 is that wild-type paddy rice (WT) and T2 are for the phenotypic map of antisense SDG725 trans-genetic hybrid rice (sdg725).

Fig. 3 is the fluorescence quantitative PCR detection result of SDG725 genetic expression in wild-type paddy rice (WT) and the antisense SDG725 trans-genetic hybrid rice (sdg725).

Embodiment

Method among the following embodiment if no special instructions, is ordinary method.The primer is finished by Ying Jun Bioisystech Co., Ltd, and examining order is finished by Shanghai Sani's bio tech ltd.

The acquisition of embodiment 1. paddy rice histone-lysine methyltransferase gene SDG725

To the gene order of the SDG8 of the Arabidopis thaliana announced among the GenBank (GenBank number: NM_106379) pass through sequence alignment, from rice genome, obtain homologous sequence, and according to 5 ' and 3 ' end sequence of this homologous sequence design pair of primers, primer sequence is respectively: 5 '-ATGGAGGAGCCTGACGGGGA-3 ', and 5 '-TCAAAATCTTTGATTATTGTTAGGA-3 '.

Extract the total RNA (Promega of paddy rice etiolated seedling, SV total RNA isolation system), take total RNA of paddy rice as template, with the synthetic cDNA (user manual according to Plant RT-PCR Kit 2.01 (TaKaRa) carries out) of AMV ThermoScript II (TaKaRa).Take cDNA as template, the full length cDNA sequence of pcr amplification paddy rice histone-lysine methyltransferase gene SDG725.50ul PCR reaction system comprises: template 2ul, high-fidelity enzyme KOD plus (TOYOBO) 1ul, 10 * damping fluid 5ul, 2.5uM dNTP 8ul, 5 of 20uM ' and each 1ul of 3 ' primer, water 32ul.Reaction conditions is: 94 ℃ of denaturations 2 minutes; 94 ℃ of sex change 30 seconds, 55 ℃ of annealing 1 minute, 68 ℃ were extended totally 30 circulations 6 minutes and 30 seconds.After reaction finishes, the PCR product is carried out 0.8% agarose gel electrophoresis to be detected, recovery and purifying be the amplified fragments of 6500bp approximately, it is cloned among the carrier pUC19 (TaKaRa), obtain containing the recombinant plasmid that reclaims fragment, show that through order-checking paddy rice histone-lysine methyltransferase gene SDG725 full-length cDNA has the nucleotide sequence of the SEQ ID No:1 in the sequence table, SEQ ID No:1 in the sequence table is by 6453 based compositions, its encoder block is from 5 ' end 1-6450 bit base, coding has the protein of the amino acid residue sequence of the SEQ ID No:2 in the sequence table, proteins encoded called after SDG725.

The protein-active of embodiment 2. paddy rice histone-lysine methyltransferase gene SDG725 is measured

The pUC19-SDG725 that obtains take embodiment 1 is as template, pcr amplification SDG725 is from the nucleotide sequence of 5 ' end 3646-4656 position (comprise the histone-lysine methyltransferase active centre---SET structural domain), 5 ' be respectively with 3 ' primer: 5 '-acggatccATGAATAAACAGAGTGATCTTATTC-3 ', and 5 '-actcgagTCAACCACCGATGTAACCCCGG-3 '.In 50 μ l pcr amplification systems, comprise masterplate pUC19-SDG725200ng, 5 ' and each 20pmol of 3 ' primer, 10Xbuffer 5 μ l, each 20 μ M of dNTP, high-fidelity KOD plus (TOYOBO) 1ul.Increase by following condition: 94 ℃ of denaturations 2 minutes; 30 seconds, 68 ℃ extensions of 30 seconds, 55 ℃ annealing of 94 ℃ of sex change 1 minute, totally 30 circulations.The PCR product is cloned into coli expression carrier pGEX-4T1 (GE life sciences) through BamH I and XhoI restriction enzyme site, and it obtains colibacillus expression plasmid pGEX-4T 1-SDG725set after correct sequence verification.

The SDG725set recombinant protein Expression in Escherichia coli purifying that GST merges carries out according to the user manual of Glutathione Sepharose 4Fast Flow (GE life sciences).

The active determination in vitro of ZNFN3A1 is pressed the (Nature such as Rea, 2000, method 406:593-599) is carried out: survey the damping fluid (50mMTris-HCl (pH 8.5) that lives at 30 μ l MAB, 20mM KCl, 10mM MgCl2,250mM Sucrose, 100 μ M ZnCl2, among the 10mM β-mercaptoethanol), the enzyme (GST-SDG725set) that adds, substrate (core histones H2A, H2B, H3 and H4 are available from Millipore) and 250nCimethyl-14C-SAM (GE life sciences), mix, 37 ℃ were reacted 1 hour.Reaction product is separated and dyeing-decolorzing through 15%SDS-PAGE, gel radioautograph behind 70 ℃ of vacuum-drying 2hrs.

Fig. 1 middle and upper part is the SDS-PAGE electrophoresis result, and the bottom is the radioautograph result, and GST albumen is as negative control, and positive control is the methyltransgerase of a known histone H 3.The result shows that GST-SDG725set has the methyl transferase activity of histone H 3.

Embodiment 3. detects SDG725 to the regulating and controlling effect of rice growth

One, the structure of SDG725 antisense expression vector

Utilize RNA to disturb (RNA interference, RNAi) technology, take the SDG725 gene as target gene, make up rna interference vector.The pUC19-SDG725 that obtains take embodiment 1 is as template, and is with the nucleotide sequence of two couples of different primer PCRs amplification SDG725 from 5 ' end 3639-3745 position, double-stranded as the complementary DNA of hairpin structure respectively.Used primer pair 1 is: 5 '-aaggatccACCTCGCATGAATAAACAGAG-3 ' and 5 '-aaaagcttCAGTTTTGCAAGCAACAACTG-3 ', gained PCR product is fSDG725i (forward).Primer pair 2 is 5 '-aagaattcCAGTTTTGCAAGCAACAACTG-3 ' and 5 '-aaactagtACCTCGCATGAATAAACAGAG-3 ', and gained PCR product is rSDG725i (reversed).For the ease of follow-up vector construction, added respectively the restriction enzyme site of BamH I and HindIII at 5 ' and the 3 ' end of fSDG725i, introduced respectively the restriction enzyme site of EcoRI and SpeI at 5 ' and the 3 ' end of rSDG725i.

For the ease of the RNAi Vector construction, introduced respectively the restriction enzyme site of HindIII and EcoR I by PCR method the PDK intron of positive and negative two dna moleculars (being provided by French molecular biology of plants institute) two ends are provided, the primer is: 5 '-aaaagcttCCAATTTGGTAAGGAAATAATT-3 ' and 5 '-aagaattcTTTCGAACCCAGCTTCCC-3 '.

With fSDG725i through BamH I and HindIII double digestion, the PDK intron is through HindIII and EcoR I double digestion, rSDG725i is through EcoR I and SpeI, connect in the pHB carrier (being given by woods letter a surname seminar of Shanghai plant physiology institute of the Chinese Academy of Sciences) through BamH I and XbaI double digestion, namely get rna interference vector pHB-fSDG725i-PDK intron-rSDG725i.

Two, SDG725 antisense expression vector rice transformation plant

With reference to (Plant Mol Biol, 1997, method rice transformations 35:205-218) such as Hiei.Plasmid pHB-fSDG725i-PDK intron-rSDG725i is changed among the agrobacterium tumefaciens EH105 by electrization, obtain positive colony through screening.The Agrobacterium EH105 that carries plasmid pHB-fSDG725i-PDK intron-rSDG725i is inoculated in the YEB liquid nutrient medium that 5ml contains the 100mg/L kantlex 28 ℃ shakes bacterium and be cultured to logarithmic phase late period, 1: 100 enlarged culturing is about 0.5 to OD600 again.Collection Agrobacterium thalline also is resuspended in the transfection media.Contaminate the fine callus of paddy rice Japan according to ordinary method, differentiation obtains the positive seedling of hygromycin resistance on coculture infection, the screening of resistance culture base and division culture medium.When treating that seedling grows to the 10cm left and right sides, seedling is moved to the plantation of outdoor solarium, sowing namely gets T1 for seed.

In order further to determine the genetic stability of this transgenosis proterties, will obtain afterwards to such an extent that T2 plants in outdoor solarium for seed, the result as shown in Figure 2, it is short and small that the paddy rice that turns rna interference vector shows obvious growth.

Three, the expression of SDG725 in the fluorescence quantitative PCR detection antisense SDG725 trans-genetic hybrid rice

Under the same conditions antisense SDG725 trans-genetic hybrid rice and wild-type paddy rice are cultivated.Press same procedure extraction rice total RNA and the synthetic cDNA of reverse transcription among the embodiment 1.Then use the expression of SDG725 in fluorescence quantitative PCR detection antisense SDG725 trans-genetic hybrid rice and the wild-type paddy rice.Detecting the SDG725 the primer is: 5 '-CTTCTGCCATACATGATGTTG-3 ', 5 '-AGAGCAATTGGCAAATGTCT-3 '.The result as shown in Figure 3, with respect to the wild-type paddy rice, the expression of SDG725 presents obvious decline in antisense SDG725 trans-genetic hybrid rice, the phenotype defective that further proves antisense SDG725 trans-genetic hybrid rice is to cause owing to the expression of SDG725 descends.

Sequence table

<160>2

<210>1

<211>6453

<212>DNA

＜213〉Oryza paddy rice (Oryza sativa ssp.japonica)

<400>1

atggaggagc ctgacgggga agcgcgtggg cgggaggatc atgctgcggt tgggcggttg 60

ggaggagagg agggtgctgt tggcggcggt ggcctggctt tgctcgctgt tcctgagatt 120

ggcggtgagc tcggtgatgg tgggaaggtc tgcggtggtc aggagaggcg cctgcccaca 180

gaggaagatg gagtccgaga taatggaggt ggctccgctg cagaattggt ggaatctgct 240

gtgaatgttt ctaccccttt tgaaggaagg ggtcagattg gtggtgagaa ggagtctagt 300

atgcaggagg ggtctatgaa catggcggga gagaagcatg gcagttatca tgtggaatct 360

gctgaaccca gcaatttgca gacgtgtcat gcgcccaatg gcggggtatc gaataagaca 420

ttatttgctc ccttcagcga agttttctcc agtgataaca gccatatgcg ttacttgctg 480

gacaaagcga cagaggggag catttgtgag catggtgatt tggcggacag taaagatgat 540

ttgggtggcg caacggacgt aaagacaaac acagaagatt tacagatggt ttgcacgaaa 600

ccacattgtg atagcgaggg tttgtcggat ttgcataatg acagtgagcg atggccacaa 660

gtggttgatg gggtgggatt tacgataaag ggtaataatg agctgaaaca agttgatttg 720

ataccaaaaa ttgaagctga agtctctagg tcggtggagg atgattcaat cccttctttt 780

tctggtggta ttgatgattc tttacgcaag gcaggttgtg cgtgtgaaac tctcaatgat 840

atggggatgt ctcatatggc caatggtgat ttgtggtgca atgttttata tgcccctctt 900

agtgaaggat gccagtccaa ggatgctcga cacatagcag tcatggggaa caaggtgaca 960

caggggagtc aatgcgggca gggtgatttg gcgtgtgatg gaattgtttt gcgaggtggg 1020

gtagatgtag agaaaagtct agatgattta cagatgtgtt ccaaggaacc acaatgtgat 1080

aacaagggct ttccatattt gacagaattt ggtgtccagc agccatcata tggcatgaat 1140

gtcatatgtt cgaagacaga tcctaaccat cagctggaaa aggatgaatt gttgacaaat 1200

actagaggag agttctctag ttctatccat gaggattcag ttccttcaat ttctgtgagc 1260

tctgttgatt ttacttttga tggcaatgct ggtcagattg gtaaaacatc tgagcataga 1320

gcaatcatgg agaaagtgtc gcatggttca cagcgaggag gtgtactttc ctgtgagagt 1380

aggtctttaa aggagtctca tgcagatgag aatcagagtt ccacattgga ggttaagaca 1440

tgtgaagagg gcttacagac aggtcaagtg gaaccatgcc acagcatcgt agctttgtcg 1500

gattcaggga agtatggcac cgacatatta ccacgtggtg gtgacggtct gagatcgatg 1560

actggtgcta accatgagct ggtaaaggat gattttcatc caaaaagtga tgtagtggtc 1620

tcctgtccag tggatgaggc atcgattccc tctaactata atagccccat tgatgttctt 1680

ttatacaaag aggatggttt agttggtgag atatctgaaa atagaattgg tgtggagaaa 1740

ttggctcatg atttgctcgg agaagttatg ctttcatttg acagcaggcc tcagactgag 1800

gcttctggag atgagaatca acacttctgg atggatgttc caaagggttc aactgcatct 1860

gtttgtgaag tagaaaatac aggcactaga agatcttgtg atccctgtgc tgaaatagag 1920

tttccactcc aacaaagtcg tgaaaagcat gtgatctctg aatccccccc agagagagat 1980

ctgactagtt cgtcccataa cctaccttgt gaaaatgaac cttgttatag tggcagggaa 2040

acacccgcct tctgcctagg ccatcaagat tctgctggtc ttggactgga atcttcagac 2100

tgtttggtac aagagctcaa tacgtgtact tccactgatg acaaagcttg ctctgttgat 2160

tttgttgaga atggtaatgg ttctcataac caaaaggaag tgccggtgat tttcttcagg 2220

cggaggaatc cagtaagagc tgcctcttca agaaattcta attttgagaa gtgtgaccag 2280

ataaacaaat caggtaatag tacacgcaaa tctaagaagg ttgacagtgt aagctcatta 2340

cttaaaagca ccatgattaa gttcccaaac aaaaccacaa agggaagaag tggcatcaat 2400

aggccattga actcttctgc ttggggcagc ctacaaaagc taatggatgg tttcaatcag 2460

aactgtggtc cttcaacttc tcgttctcat caaacttgtt tgggaaaaga aatatcaaat 2520

agaggatcta gcgagaaaaa acagctatct attcggaaaa ttcgaacttc aagatgttca 2580

aaatataaaa acacatcact ttctgatatt ggatatttag caggtgaatt gaatggccaa 2640

ccgacctgtt cagtgaggat tgatactaat gtttcttctg atgcattgtt caattctccg 2700

aatggtgctc acaaagctgc acagtgtgtt gaaggtaatc atactctaaa attaacatct 2760

agcctgactg acacacaaca gtttggcttg gagaatgtta ctcaagaaac atgccctggg 2820

tacatccatg gagagtgtgg tacttcaact tctgaacgtt ctctaaataa tatagttggg 2880

ttttcaccag actctgtttt ggatatagct tctgttacat gtgaaagcaa cacttctgca 2940

acccttgatg ttatagtgca tgaaaaccca tcttgtcctg gtggattgat tggaggtggt 3000

cttcgtgcat ctgctttatc tacttctcac tgtgaaaatc atcatgcttc atcattgatg 3060

gatttggagc agcaggtcaa aactgtgagg gagaacgaca tgggagagga agatgtcatt 3120

ccatcacatg ccatgatgta caatgatatt ggtgaaggaa agcaaacttt agcgaagtcc 3180

aatacgat gaggaaaggtag aaatgtggga aagcaggaat gccgaaagaa agatggaaag 3240

aagggaaaaa acataaacaa aaatagaagt tccaccaaaa tttcatctag tgaagcttca 3300

aaactcgtgt ccttttctaa tgattcacct tcacttgatc catctgagtt gctgcttcat 3360

acgagacctc caaagtttgg ttcttgttct aaggtcgtaa cttctgccat acatgatgtt 3420

ggtatgcatg gatatgacaa tatgcgtcct tttggaattg acaatgatga cgaagggagt 3480

gcatttgaca atgtgaaatc actaaggcgc aagaaaaagg atagtcatgg aggaaagaag 3540

ggtaaggtgc gggatccaca tgggaagggc agaagcaaga agaaaaatat agctgataac 3600

acctacggtt tgccggctca gttaactgac ctgtcagaac ctcgcatgaa taaacagagt 3660

gatcttattc ctgctgctga acttgtattc aagaactctt ctgccgtatc tgttgaatta 3720

cctgcagttg ttgcttgcaa aactgatggt gcatctgtac caccagcacc tgcttgggtt 3780

tgttgtgatg attgcgaaaa atggcgctgc ataccaactg aactggcaga taaaatctca 3840

aaagaaaatc tcagatggac ttgtaaggaa aacgaagata agacatttgc caattgctct 3900

ataccacaag agaagacaga tgatgagatc aatgcagagc ttgggctttc agatgcttct 3960

gctgatgaag ctaatggcga tggatcaaac tcgaaagctt ctggagaacc aaactttgca 4020

cttctcaggt caaacttgtt tctacatcgt aaccgcagga cacaatccat tgatgagagc 4080

atggtatgca attgtaagcc gcctcacgat gaccgaatgg gttgtagaga tggttgcttg 4140

aacaggatac tcaacattga atgcaccaaa cgtacatgtc catgtgggga gcactgttcc 4200

aatcagcagt tccaaaggcg cacctatgca aaacttggta agttccatac tggtaaaaag 4260

ggctatggat tgcaattgaa ggaagatgta tctgaaggac gattcctcat tgaatatgtt 4320

ggagaggtcc ttgatataac ggcttatgaa tcccgccaaa ggtattatgc ctctaaaggc 4380

cagaagcatt tctatttcat ggcacttaat ggtggtgagg tgatagatgc ttgtactaaa 4440

ggaaacttgg gccggttcat caatcatagc tgcagtccta attgccgtac agagaagtgg 4500

atggtcaatg gtgaagtctg cattggaata tttgctatga ggaacatcaa gaagggtgaa 4560

gaattgacgt ttgattacaa ctatgttcgt gtatctggtg ctgctcctca gaaatgcttt 4620

tgcggtactg ccaaatgccg gggttacatc ggtggtgaca tatcaggtgc tgatatgatt 4680

actcaagatg atgctgaagc agggactttc gaacctatgg ctgttcagga ggatgctgag 4740

gaagtacttg gtgcaaatgg tttgtcctct catggcacac atctagatat tgtcgaccat 4800

gaagcttcca ctaaaacaga agattcaaat gattgcccat ctgtgaaccc accagagtta 4860

gaatctgagc aacaaacttc aggaacctta tttgacacaa gtgagccaga aaattcctta 4920

gaagcattga gcccacagga tgatgaagat gtcgtccgca cacctgtcca tgtgtcccgg 4980

acagttgaga gtacgtcgcg gcagtttcca gaatatggta ctcggtcatc agaaattttg 5040

caaagggctc catgcacact ggatggacca aaggttccaa gcacaacaaa tggaattccg 5100

cctagttccg atttggggag ccactgggta ccaggtttcc acgctaataa gaaaaccaat 5160

gtaaaacatc atttgattct gaatccatca tcagctccta ttgacagtga gcacattttg 5220

ggagttgaag gaagattgaa cagcttgctt gatgtaaatg gaggtattag caaacgaaaa 5280

gacgcaacaa atggatactt gaagcttctc cttgtgactg cagcggaagg tgacaatgct 5340

gggggcacgt ctaaaagtgt aagggatctt tcgttaattc ttgatgcact tctgaaaaca 5400

agatccaatt ctgtcctgtt ggatatcatc aataagaatg gactgcaaat gcttcacaat 5460

atattaaagc agaataaaag cgatttccat aggataccta taataagaaa gcttgtgaag 5520

gtacttgagt ttctagcttc caagggcatt ctgacatctg aacatataaa tggtggtcct 5580

cgatgtgctg ggacagaaag cttcagggag tcgatgttgg gcttgttaag gcataatgat 5640

atgcaggttc aacagattgc tcggaacttt cgtgatagat ggatccaatg ggcccctcga 5700

aatatttcaa gaaatgagcc aacagaatat tcacgtgcat ccatatctgc acacgatatt 5760

catgtcattt caacagctgg tggatctttt ccaacttcag ccaacactat ggattggaaa 5820

tccatcagaa gaaaacgcaa gagccgttgg gattaccaac cagatgacca ctataaaatg 5880

ggtggcctga aaattcagaa agtttgtcct gtacaaagtg agtttcggac tggctcggtg 5940

ggaaataagt tgcatggtaa ttggggaacg aacagctccc acaatgatgt tcctgtggtg 6000

ggaagttcag ccgatggtgc agatgatgaa gcacctcctg gatttgagtc tcagcaggaa 6060

agtcggcctg gacaagcttg tttagaatcg ggggtttctc caggattata cttggagagg 6120

tatcaacata acttgactat ttcatacggg attccaatcg cttttgttga gcattttgga 6180

accccagaag ttgaaggagg gccatgtcgc aaaaactgga aagtcgcacc tggtgtgcct 6240

ttccagccct ttccaccatt gccgccctat ccaagaggga gtccttgtcc ttccactcag 6300

atgtctcagc atgaacacaa cagtttggga cattgtggca gggctgcaaa tagagacggg 6360

agaatacata ggaattggag aaatggggcg agaacaaaat ttccatacaa tcatcaagga 6420

cggagatttc ctaacaataa tcaaagattt tga 6453

<210>2

<211>2150

<212>PRT

＜213〉Oryza paddy rice (Oryza sativa ssp.japonica)

<400>1

MEEPDGEARG REDHAAVGRL GGEEGAVGGG GLALLAVPEI GGELGDGGKV CGGQERRLPT 60

EEDGVRDNGG GSAAELVESA VNVSTPFEGR GQIGGEKESS MQEGSMNMAG EKHGSYHVES 120

AEPSNLQTCH APNGGVSNKT LFAPFSEVFS SDNSHMRYLL DKATEGSICE HGDLADSKDD 180

LGGATDVKTN TEDLQMVCTK PHCDSEGLSD LHNDSERWPQ VVDGVGFTIK GNNELKQVDL 240

IPKIEAEVSR SVEDDSIPSF SGGIDDSLRK AGCACETLND MGMSHMANGD LWCNVLYAPL 300

SEGCQSKDAR HIAVMGNKVT QGSQCGQGDL ACDGIVLRGG VDVEKSLDDL QMCSKEPQCD 360

NKGFPYLTEF GVQQPSYGMN VICSKTDPNH QLEKDELLTN TRGEFSSSIH EDSVPSISVS 420

SVDFTFDGNA GQIGKTSEHR AIMEKVSHGS QRGGVLSCES RSLKESHADE NQSSTLEVKT 480

CEEGLQTGQV EPCHSIVALS DSGKYGTDIL PRGGDGLRSM TGANHELVKD DFHPKSDVVV 540

SCPVDEASIP SNYNSPIDVL LYKEDGLVGE ISENRIGVEK LAHDLLGEVM LSFDSRPQTE 600

ASGDENQHFW MDVPKGSTAS VCEVENTGTR RSCDPCAEIE FPLQQSREKH VISESPPERD 660

LTSSSHNLPC ENEPCYSGRE TPAFCLGHQD SAGLGLESSD CLVQELNTCT STDDKACSVD 720

FVENGNGSHN QKEVPVIFFR RRNPVRAASS RNSNFEKCDQ INKSGNSTRK SKKVDSVSSL 780

LKSTMIKFPN KTTKGRSGIN RPLNSSAWGS LQKLMDGFNQ NCGPSTSRSH QTCLGKEISN 840

RGSSEKKQLS IRKIRTSRCS KYKNTSLSDI GYLAGELNGQ PTCSVRIDTN VSSDALFNSP 900

NGAHKAAQCV EGNHTLKLTS SLTDTQQFGL ENVTQETCPG YIHGECGTST SERSLNNIVG 960

FSPDSVLDIA SVTCESNTSA TLDVIVHENP SCPGGLIGGG LRASALSTSH CENHHASSLM 1020

DLEQQVKTVR ENDMGEEDVI PSHAMMYNDI GEGKQTLAKS NTMRKGRNVG KQECRKKDGK 1080

KGKNINKNRS STKISSSEAS KLVSFSNDSP SLDPSELLLH TRPPKFGSCS KVVTSAIHDV 1140

GMHGYDNMRP FGIDNDDEGS AFDNVKSLRR KKKDSHGGKK GKVRDPHGKG RSKKKNIADN 1200

TYGLPAQLTD LSEPRMNKQS DLIPAAELVF KNSSAVSVEL PAVVACKTDG ASVPPAPAWV 1260

CCDDCEKWRC IPTELADKIS KENLRWTCKE NEDKTFANCS IPQEKTDDEI NAELGLSDAS 1320

ADEANGDGSN SKASGEPNFA LLRSNLFLHR NRRTQSIDES MVCNCKPPHD DRMGCRDGCL 1380

NRILNIECTK RTCPCGEHCS NQQFQRRTYA KLGKFHTGKK GYGLQLKEDV SEGRFLIEYV 1440

GEVLDITAYE SRQRYYASKG QKHFYFMALN GGEVIDACTK GNLGRFINHS CSPNCRTEKW 1500

MVNGEVCIGI FAMRNIKKGE ELTFDYNYVR VSGAAPQKCF CGTAKCRGYI GGDISGADMI 1560

TQDDAEAGTF EPMAVQEDAE EVLGANGLSS HGTHLDIVDH EASTKTEDSN DCPSVNPPEL 1620

ESEQQTSGTL FDTSEPENSL EALSPQDDED VVRTPVHVSR TVESTSRQFP EYGTRSSEIL 1680

QRAPCTLDGP KVPSTTNGIP PSSDLGSHWV PGFHANKKTN VKHHLILNPS SAPIDSEHIL 1740

GVEGRLNSLL DVNGGISKRK DATNGYLKLL LVTAAEGDNA GGTSKSVRDL SLILDALLKT 1800

RSNSVLLDII NKNGLQMLHN ILKQNKSDFH RIPIIRKLVK VLEFLASKGI LTSEHINGGP 1860

RCAGTESFRE SMLGLLRHND MQVQQIARNF RDRWIQWAPR NISRNEPTEY SRASISAHDI 1920

HVISTAGGSF PTSANTMDWK SIRRKRKSRW DYQPDDHYKM GGLKIQKVCP VQSEFRTGSV 1980

GNKLHGNWGT NSSHNDVPVV GSSADGADDE APPGFESQQE SRPGQACLES GVSPGLYLER 2040

YQHNLTISYG IPIAFVEHFG TPEVEGGPCR KNWKVAPGVP FQPFPPLPPY PRGSPCPSTQ 2100

MSQHEHNSLG HCGRAANRDG RIHRNWRNGA RTKFPYNHQG RRFPNNNQRF 2150

Claims (6)

1. a paddy rice histone-lysine methyltransferase is characterized in that amino acid residue sequence is shown in SEQ ID No:2.

2. the gene of a paddy rice histone-lysine methyltransferase as claimed in claim 1 is characterized in that nucleotides sequence classifies SEQ ID No:1 as.

3. the expression vector that contains paddy rice histone-lysine methyltransferase gene claimed in claim 2.

4. the engineering bacteria that contains paddy rice histone-lysine methyltransferase gene claimed in claim 2.

5. the application of paddy rice histone-lysine methyltransferase gene claimed in claim 2 in adjusting and controlling rice grows.

6. application according to claim 5 is characterized in that: with the antisense expression vector rice transformation of described paddy rice histone-lysine methyltransferase gene, obtain the short and small paddy rice of plant strain growth.

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