CN1295334C - Wheat antidisense related gene TaEDR1 and its application - Google Patents

Wheat antidisense related gene TaEDR1 and its application Download PDF

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CN1295334C
CN1295334C CNB2005100178062A CN200510017806A CN1295334C CN 1295334 C CN1295334 C CN 1295334C CN B2005100178062 A CNB2005100178062 A CN B2005100178062A CN 200510017806 A CN200510017806 A CN 200510017806A CN 1295334 C CN1295334 C CN 1295334C
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gene
wheat
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leu
taedr1
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CN1757730A (en
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牛吉山
郭天财
张丽娜
王映红
洪德峰
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Henan Agricultural University
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Henan Agricultural University
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Abstract

The present invention relates to the technical field of plant gene engineering, particularly to a gene TaEDR1 relevant to disease resistance of wheat and an application thereof. The gene TaEDR1 relevant to disease resistance of wheat is separated from new strain 99-2439 of disease resistance of wheat having high resistance degree to powdery mildew, the full length of cDNA of the gene is 3050 bp, and the gene comprises a 5'-non-coding region in the length of 70 bp, a 3'-non-coding region in the length of 70 bp, a coding region in the length of 2877 bp, a terminator and a poly-A tail in the length of 30 bp, wherein the 3'-non-coding region comprises one typical tailing signal sequence. The coding protein of the gene is a mitogen protein kinase whose the full length is composed of 959 amino acids. The gene TaEDR1 relevant to disease resistance of wheat can be applied to disease resistance improvement of plants, new disease resistance relevant gene synthesis and wheat gene chip manufacture, and has high practical application value.

Description

Wheat antidisense related gene TaEDR 1 and application thereof
One. technical field: the present invention relates to the plant gene engineering technology field, particularly relate to a kind of wheat antidisense related gene TaEDR 1 and application thereof.
Two. background technology:
Plant transgene research starts from phase early 1980s, nineteen eighty-three Zambryski has obtained the first routine transfer-gen plant in the world, Horch had created the Ye Panfa in the Agrobacterium-mediated Transformation in 1985, henceforth, set up agrobacterium-mediated transformation respectively, virus-mediated method, the PEG mediated method, electricity swashs perforation method, microinjection, pollen tube passage method, supersonic method, particle bombardment etc., the species of transgenosis success constantly enlarge, relate to more than 50 species totally 110 various plants, wherein, the transgenic plant that agrobacterium-mediated transformation obtains account for more than 85% of transgenic plant sum, the transgenic plant that particle bombardment obtains account for about 10%, dicotyledons is the natural host of Agrobacterium, utilize Agrobacterium Ti-plasmids mediated method to set up the gene transfer system of multiple dicotyledons, and some good foreign genes have been changed over to dicotyledons, bred transformed variety, but utilized agrobacterium-mediated transformation relatively slow the work progress that monocotyledons carries out gene transformation.
Wheat is one of topmost food crop, but its engineered flow of research has lagged behind other crop, and wheat transgenic research starts from the initial stage eighties.Vasil in 1992 etc. utilize the particle gun mediated method to obtain the first routine wheat transgenic plant in the world, and once monarch's happiness, Cheng Zhuomin etc. utilized pollen tube passage method to obtain the wheat transgenic plant respectively in 1994.In afterwards several years, wheat transgenic is studied basically by means of the particle gun mediated method.According to statistics, up to the present obtain in the report of wheat transgenic, particle bombardment accounts for about 90%, and other method only accounts for about in the of 10%, and reason is the method comparative maturity of particle gun mediated method, and agrobacterium-mediated transformation is also relatively more difficult.But, compare with the particle gun mediated method, that agrobacterium-mediated transformation has is simple to operate, cost is low, transformation efficiency is high, good reproducibility, can import advantages such as large fragment DNA, and the gene that imports is generally single copy and integrates, be unlikely to the reticent phenomenon of producer, yet the agriculture bacillus mediated transgenosis of wheat is an a difficult problem in the world always, though Hess, Mooney etc. did trial once, fail to obtain transfer-gen plant.Raising along with gene isolation technology and clone's means, to there be more important gene relevant to be cloned out from now on the wheat improvement, some practical problemss in the while Wheat Production, come out in succession as aphid, head blight, Powdery Mildew, rust, gaeumannomyces graminis disease, banded sclerotial blight, soil-borne disease viral disease, arid, problem such as saline and alkaline, and genetic engineering breeding may be best solution route.
Plant is subjected to the infringement of many pathogens in the process of growth, the phytopathy original of a great variety comprises virus, bacterium, mould and nematode etc.The pathogen invaded plants causes two kinds of results: 1. breeding in host plant of pathogen success causes relevant illness; 2. host plant produces defense response, kills pathogen or stops its growth.Disease is the major obstacle of Wheat Production, and most of diseases of wheat belong to fungal disease, and it is one of approach of control wheat diseases that the external source anti-fungal gene is imported wheat, and agrobacterium-mediated transformation generally commonly used carries out gene transformation to wheat.Number of patent application is 03109435.X, denomination of invention is for disclosing the agriculture bacillus mediated method of wheat being carried out gene transformation of a kind of usefulness in " the agriculture bacillus mediated method of wheat being carried out gene transformation of a kind of usefulness ", it is that WHEAT CALLUS and the Agrobacterium that contains the Ti-plasmids carrier are cultivated altogether, foreign gene on the plasmid vector is transferred in the acceptor gene group, but before conversion, will be carried out resuspended with the nutrient solution that contains ethyl sulfonic acid, Methionin, octopine, glutamine, Syringylethanone, glucose, maltose etc.At present worldwide, fewer comparatively speaking to the research of separation, clone and the disease resistance response of wheat resistance genes and disease-resistant wheat genes involved.
Three. summary of the invention:
The objective of the invention is: overcome the deficiency that exists in the present disease-resistant wheat genes involved research process, a kind of new wheat antidisense related gene TaEDR 1 and application thereof are provided.
Technical scheme of the present invention is:
A kind of wheat antidisense related gene TaEDR 1 is characterized in that: the full length cDNA sequence of TaEDR1 gene, the based composition of its nucleotide sequence: 896a 630c 758g 766t
1 cgccaaactg?aaataaccaa?gagaggaaat?attttcgaaa?cccaaaccct?aggtgctcgt
61 ggggagcgcc?atgaagatcc?cgtttgtgac?caagtggtcg?caccgatcca?gcgagcccgc
121?ggggccgtcg?aattcggctg?cagcgcagca?gcagcagcag?cagcaggcgc?cgtctcctcc
181?tcctcccgtg?gcgtcgacag?aggcggcagg?ggatgagttc?attctgcagg?aggaagagta
241?ccagatgcaa?ctggcgttgg?cgctatcagc?gtcggcgtcg?ggcgccgagg?gcgctgggga
301?tcccgacggg?gagcagatca?gaaaggcgaa?gctgatgagc?ctcgggaagg?gccacccggt
361?caccaacagc?gatcgtggcg?ggggagacac?cccggagtcg?ctctcccgcc?gttacaggga
421?ctataacttt?cttgattaca?atgagaaagt?aattgatgga?ttctacgacg?tatttggcct
481 ctctgcggga?tcatctgggc?agggcaaaat?accttcactg?gcagagcttc?agatgagcat
541 tggggatctt?ggatatgaag?taattgtggt?tgactataaa?tttgataatg?ctctgcagga
601 gatgaaggaa?gtagcagaat?gctgcctgtt?gggctgtcct?gacattacag?tattggtgcg
661 acgaatagct?gaagttgttg?cagatcacat?gggtggtcca?gtgatcgatg?caaatgaaat
721 gatcactagg?tggttgagca?aaagcattga?gcagaggaca?tcacaccaga?caagccttct
781 gcatattggc?agtatagaga?taggcttgtc?tcgccatcgt?gccttacttt?tcaagattct
841 tgctgatatt?gttggtatcc?cttgcaagct?ggttaaaggg?agtcattaca?ctggtgttga
901 agatgacgct?attaacataa?taaagatgga?tgacaaaagg?gagtttttgg?tggatgttat
961 ggctgctcca?gggactctca?ttccagcaga?tgtctttaat?tcaaagggta?ctccattcaa
1021?cttcagtcaa?acattgggtc?agaatcaggt?ggtggagtca?gcaagtaaca?tcgaagatga
1081?cccagttgca?ttacagtcag?agcataaacg?taaccaaggg?catatgtttg?ccaataataa
1141?tcggatctca?gtcaatctat?caagctatga?gaatacaatg?accgctggaa?gtagtgctag
1201?tgaacctggg?acattggacc?ctaggatgca?attaggtaaa?acatcaactt?tgcctagtgc
1261?tccttccaag?cagaagaaga?atctgcaatt?gattacagac?tctcatgaaa?ctgaagagtc
1321?ccgaaaacta?tttgtggagt?tagatccttt?caatgctatt?gaatctggga?aaagctcatt
1381?ggcattcaag?ggattaaata?atagaaacaa?tgaattccaa?aggcgtagag?agaatgtagt
1441?cccaccatct?gtaagatctc?aacagccatt?ggtgatgaaa?aactggtctg?cttgcaatga
1501?catttccaac?aacaagcaat?acaatgttgc?tgatgggtca?gttcctcgga?gaaatgccac
1561?tgacaatgca?tcgtcatctc?agttggcgtt?gtcaactgca?aagcattaca?attccaatgt
1621?tagagagcta?aacgatagag?tgtatgcagc?acctgctcgt?aattatgaca?acaagatagt
1681?tggtacctcg?gctatggcca?aagcattgac?tggagagtgc?cctgacagat?cacaggtgcc
1741?acctggtctt?tattatgaca?agatgcttgg?tacctcttct?atgaatgcag?cttctacatc
1801?cggaatcggg?aaagttgcag?aaaaggaccc?tcataatgat?ccgggaaaag?gtcccatcta
1861?ttctagattt?gatggtgaac?tttctaaaaa?tgctcaagga?tttactcccg?aaagggatga
1921?gcacaaggaa?aattgtggca?gtcatgacca?caaaatgtta?tatcctgatc?caagaaagtc
1981?ccctcttgac?agattcatgg?acaggccaag?gcagagcata?gaatgtgttt?ttccatccca
2041?agttggatca?aataaggctg?acatggtgtt?ggatgaagtg?tctgaatgtg?aaatcctttg
2101?ggaagatctt?gtaatcgatg?aaagaattgg?cataggttca?tatggagaag?tctaccatgc
2161?tgattggaat?ggaactgaag?tagctgtaaa?gaagttcttg?gatcaagagt?tctatggtga
2221?tgctttagag?gaatttcgtt?gtgaagtgag?gattatgcgt?cggctccgtc?atccaaatat
2281?tgttctcttt?atgggtgcag?taacacggcc?tccacactta?tctattgtat?cagaatatct
2341?tccaagggga?agcttatata?agatcattca?tcgccctaat?tgccaaatcg?acgagaagcg
2401?taggattaaa?atggcccttg?atgtggccag?aggcatgaat?tgtcttcata?ccagtgtacc
2461?aacaattgtt?caccgggatc?taaaatcacc?aaacttgctg?gttgacgata?attggactgt
2521?gaaggtctgt?gatttcggac?tttcacgtct?gaagcacagt?acatttttgt?catcaaaatc
2581?cactgccggg?actcctgagt?ggatggcacc?agaggttttg?cggaatgagc?aatccaatga
2641?gaagtgtgat?atttacagct?ttggtgttat?cctgtgggag?ctagcaacac?taagaaagcc
2701?atggcatggg?atgaaccaaa?tgcaagttgt?gggcgcagtt?ggcttccagg?accgacggct
2761?tgacattcca?aaagaagtag?atcctatagt?tgcatcaatt?atacgtgatt?gctggcagaa
2821?ggatccaaac?ttgcgtcctt?ctttcatcca?attaactagc?tacctgaaga?cattgcaaag
2881?gcttgtaatc?ccttcacatc?aggagacagc?gagcaaccat?gtaccctatg?aaatatcttt
2941?atatcggtga?accgcacatc?tctaccccgg?ggttgtcacc?caccaagtgt?gaataagcag
3001?taattatgat?tttgtcatcg?aaaaaaaaaa?aaaaaaaaaa?aaaaaaaaaa
The albumen that described wheat antidisense related gene TaEDR 1 is coded, its aminoacid sequence is:
1 MKIPFVTKWS?HRSSEPAGPS?NSAAAQQQQQ?QQAPSPPPPV?ASTEAAGDEF?ILQEEEYQMQ
61 LALALSASAS?GAEGAGDPDG?EQIRKAKLMS?LGKGHPVTNS?DRGGGDTPES?LSRRYRDYNF
121?LDYNEKVIDG?FYDVFGLSAG?SSGQGKIPSL?AELQMSIGDL?GYEVIVVDYK?FDNALQEMKE
181?VAECCLLGCP?DITVLVRRIA?EVVADHMGGP?VIDANEMITR?WLSKSIEQRT?SHQTSLLHIG
241?SIEIGLSRHR?ALLFKILADI?VGIPCKLVKG?SHYTGVEDDA?INIIKMDDKR?EFLVDVMAAP
301?GTLIPADVFN?SKGTPFNFSQ?TLGQNQVVES?ASNIEDDPVA?LQSEHKRNQG?HMFANNNRIS
361?VNLSSYENTM?TAGSSASEPG?TLDPRMQLGK?TSTLPSAPSK?QKKNLQLITD?SHETEESRKL
421?FVELDPFNAI?ESGKSSLAFK?GLNNRNNEFQ?RRRENVVPPS?VRSQQPLVMK?NWSACNDISN
481?NKQYNVADGS?VPRRNATDNA?SSSQLALSTA?KHYNSNVREL?NDRVYAAPAR?NYDNKIVGTS
541?AMAKALTGEC?PDRSQVPPGL?YYDKMLGTSS?MNAASTSGIG?KVAEKDPHND?PGKGPIYSRF
601?DGELSKNAQG?FTPERDEHKE?NCGSHDHKML?YPDPRKSPLD?RFMDRPRQSI?ECVFPSQVGS
661?NKADMVLDEV?SECEILWEDL?VIDERIGIGS?YGEVYHADWN?GTEVAVKKFL?DQEFYGDALE
721?EFRCEVRIMR?RLRHPNIVLF?MGAVTRPPHL?SIVSEYLPRG?SLYKIIHRPN?CQIDEKRRIK
781?MALDVARGMN?CLHTSVPTIV?HRDLKSPNLL?VDDNWTVKVC?DFGLSRLKHS?TFLSSKSTAG
841?TPEWMAPEVL?RNEQSNEKCD?IYSFGVILWE?LATLRKPWHG?MNQMQVVGAV?GFQDRRLDIP
901?KEVDPIVASI?IRDCWQKDPN?LRPSFIQLTS?YLKTLQRLVI?PSHQETASNH?VPYEISLYR
The albumen that described wheat antidisense related gene TaEDR 1 is coded, its genes encoding be a mitogen protein kinase kinase kinases, comprise N-terminal sequence and C-end serine/threonine protein kitase structural domain 2 major parts.
The application of wheat antidisense related gene TaEDR 1 in the plant disease-resistant improvement.
The application of wheat antidisense related gene TaEDR 1 in synthetic new disease-resistant related gene.
The application of wheat antidisense related gene TaEDR 1 in the wheat cdna chip manufacturing.
Positive beneficial effect of the present invention is:
1. wheat antidisense related gene TaEDR 1 of the present invention can carry out the transgenosis breeding for disease resistance.This gene can be inserted into downstream of different types of promoter, be built into plant expression vector, carry out the plant disease-resistant improvement by genetic engineering technique, promotor can be the promotor of constitutive expression, as cauliflower mosaic virus 35S promoter, corn ubiquitin promoter Ubi etc., the gene that is connected the composition type expression promoter downstream will be expressed at any tissue and any developmental stage of transgenic plant; Promotor can be the promotor of pathogenic bacterium inducing expression promoter or tissue specific expression also, and the gene that is connected under these promotors is only expressed when pathogenic bacteria exists or in particular organization, makes this expression of gene be subjected to more accurate control.
2. wheat antidisense related gene TaEDR 1 of the present invention can design synthetic new disease-resistant related gene.With TaEDR1 and disease-resistant relevant mitogen protein kinase kinase kinases such as EDR1, HvEDR1 more as can be known, these proteic carboxyl terminal serine/threonine protein kitase functional domain high conservatives, and have than big-difference in N-terminal structure, therefore, N-terminal structural changes decision EDR1 albuminoid and other proteic specificitys of doing mutually, by technology such as rite-directed mutagenesis or gene order reorganization, the gene that can synthetic much has new characteristic is applied in the plant disease-resistant improvement by genetic engineering technique.In addition, wheat antidisense related gene TaEDR 1 of the present invention is the gene with disease-resistant negative regulation effect, make its protein kinase function territory inactivation by rite-directed mutagenesis, or antisense rna construct, RNA disturb plant expression vector, be applied in the plant disease-resistant improvement by transgenic technology.
3. wheat antidisense related gene TaEDR 1 of the present invention can carry out the making of gene chip.The clone of this gene can be used for the wheat cdna chip manufacturing, be applied in the research such as disease-resistant wheat, according to TaEDR1 gene cDNA sequence design PCR primer, the full-length cDNA of amplification TaEDR1 gene in containing the clone of Ben Jiyin, or the strong section of gene specific in amplification coding N-end structure territory, carry out the making of gene chip.
4. wheat antidisense related gene TaEDR 1 of the present invention plays an important role in the wheat anti-powdery mildew reaction.Wheat is when the attack that is subjected to wheat powdery mildew, and the expression of wheat antidisense related gene TaEDR 1 in wheat leaf blade strengthens, and the enhancing amplitude can reach 3-5 doubly (referring to Fig. 1); Wheat antidisense related gene TaEDR 1 expression amount in wheat leaf blade is higher, in young stem and young fringe, express medium, at root expression amount lower (referring to Fig. 2), the express spectra of wheat antidisense related gene TaEDR 1 changes has proved that it plays an important role in the wheat anti-powdery mildew reaction.
5. the approach of wheat antidisense related gene TaEDR 1 of the present invention is one of important channel of disease-resistant wheat.The coded albumen of wheat antidisense related gene TaEDR 1 of the present invention is a mitogen protein kinase kinase kinases (MAPKKK), comprise N-terminal sequence and C-end serine/threonine protein kitase structural domain 2 major parts, when the attack that is subjected to wheat powdery mildew, the expression of TaEDR1 gene in wheat leaf blade strengthens, the enhancing amplitude can reach 3-5 doubly (referring to Fig. 1), TaEDR1 gene expression amount in wheat leaf blade is higher, in young stem and young fringe, express medium, at root expression amount lower (referring to Fig. 2), the express spectra of TaEDR1 changes has proved that it plays an important role in the wheat anti-powdery mildew reaction, the coded proteic aminoacid sequence of wheat antidisense related gene TaEDR 1 and the barley HvEDR1 that has cloned (the GanBank query ID: gi:11127922) have 92% identical, from comparative result as can be seen: the approach of wheat antidisense related gene TaEDR 1 of the present invention is one of important channel of disease-resistant wheat.
Four. description of drawings:
Fig. 1 is for changing by the expression of sxemiquantitative RT-PCR technical Analysis TaEDR1 gene in wheat anti-powdery mildew reaction, and M is a molecular weight standard on the figure, 0,1,12,24 etc. digital be that wheat powdery mildew is inoculated induction time; Actin is the wheat actin gene, be the stably express gene, contrast as changes in gene expression, TaMlo is a disease-resistant related gene of wheat, expression is strengthened by inducing slightly of white powder germ, and as the contrast of gene expression analysis, the result as seen, the TaEDR1 gene is expressed in white powder germ inoculation back and is strengthened, the white powder germ inoculate back 24 hours expression of gene amounts reach when not inoculating about 3-5 doubly;
Fig. 2 is for changing by the expression of sxemiquantitative RT-PCR technical Analysis TaEDR1 gene in the wheat different tissues, M is a molecular weight standard on the figure, swimming lane 1,2,3,4,5,6,7 is respectively spire, middle leaf, boot leaf, young fringe, young stem, root, white powder germ inductive root, Actin is the wheat actin gene, be the stably express gene, as the contrast of changes in gene expression.
Five. embodiment:
Embodiment one: clone's process of wheat antidisense related gene TaEDR 1:
Wheat TaEDR1 gene is to isolating the very high disease-resistant wheat new lines " 99-2439 " of Powdery Mildew resistance level.Inducing back 22 hours blade with the white powder germ is the total RNA of material extraction (Yeast Nucleic Acid), further again separating mRNA (messenger RNA(mRNA)), by reverse transcription mRNA being transcribed into cDNA (complementary DNA (cDNA)), is that template is carried out gene clone with this cDNA.
According to a pair of merger primer of Arabidopis thaliana EDR1 gene and homologue thereof design (sequence is: 5 '-GCIGTIAAIAAITTI (T/C) TIGA (T/C) CA (G/A) GA-3 ' and 5 '-GCIAAIGAIGGIC (G/T) IAI (G/A) TT IGG (G/A) TC-3 '), the cDNA fragment that has obtained to represent the 627bp of wheat TaEDR1 gene to grow by the reverse transcription PCR technology.On the basis of this sequence, designed one 5 again '-the RACE gene-specific primer (sequence is: 5 '-CGGAGCCGACGCATAATCCTCACTT-3 '), by rapid amplifying cDNA end technology obtained the long gene 5 of 2268bp '-terminal sequence.Subsequently, again 5 '-end non-coding region design gene-specific primer (sequence is: 5 '-TAGGTGCTCGTGGGGAGCG-3 '), obtained the full length cDNA sequence clone of wheat TaEDR1 gene by rapid amplifying cDNA end technology.
Concrete clone technology:
CDNA's is synthetic: use TRIzol Reagent extracts total RNA.With Poly (A) Tract MRNA separation system III isolates mRNA.Use SMART TMRACE cDNA amplification kit synthesizes cDNA.
Reverse transcription PCR: increase with high-fidelity DNA polymerase.Amplification program is: earlier 94 ℃ of sex change 2 minutes; Connect 94 ℃ of sex change of 30 round-robin 30 seconds, annealed 30 seconds for 52 ℃, 72 ℃ were extended 1 minute; Extended 7 minutes at 72 ℃ at last.
Rapid amplifying cDNA end: use special test kit (SMART TMRACE cDNA AmplificationKit) increase, solution preparation by specification carries out, and amplification program is 94 ℃ of sex change of 5 round-robin 15 seconds, anneals and extends 3 minutes for 72 ℃; 94 ℃ of sex change of 25 round-robin 15 seconds were annealed 30 seconds for 68 ℃, and 72 ℃ were extended 3 minutes; Extended 7 minutes at 72 ℃ at last.
Product reclaims and sequencing analysis: amplified production is electrophoretic separation on 1% sepharose.The sepharose that will contain target DNA fragment cuts down, and reclaims equal-volume chloroform extracting and purifying, the alcohol precipitation of 2 times of volumes with the liquid nitrogen multigelation.The dna fragmentation that reclaims is connected it with the T4-DNA ligase enzyme after dissolving with ultrapure water with pGEM-T Easy carrier, 4 ℃ of connections are spent the night.Use CaCl 2The thermal shock method 42 ℃ of thermal shocks 90 seconds, will connect product and be transformed in the intestinal bacteria DH10B bacterial strain.Extract plasmid DNA with plasmid a small amount of extraction method.Precious biotech firm checks order with the ABI377 automatic sequencer in Dalian.Sequencing result carries out homologous sequence inquiry relatively at American National bioinformation center website NCBI (http://www.ncbi.nlm.nih.gov) with relevant networking website with BLASTp computer aided program (Altschul etc. 1997) with BLASTt.Guard the functional domain data analysis in the NCBI website with the method for (2003) such as Marchler-Bauer.
In GenBank, carry out the similarity inquiry relatively, prove that the full-length cDNA of being cloned is new wheat cdna, called after TaEDR1 gene, the partial sequence of this gene is logined in GenBank as the new gene of wheat, is numbered: AY743662.
The coded proteic aminoacid sequence of wheat antidisense related gene TaEDR 1 and the barley HvEDR1 that has cloned (the GanBank query ID: gi:11127922) have 92% identical, its comparative result is as follows:
TaEDR1:1 MKIPFVTKWSHRSSEPAGPSNSAAAQQQQ----QQQAPSPPPPVASTEAAGDEFILQEEEYQMQLALALSASASGAEGAGDPDGEQIRKA 86
MKIPFVTKWSHRSSEPAGPSNSAAAQQQQ APS PPVASTEAAGDEFILQEEEYQMQLALALSASASGAEGAGDPDGEQIRKA
HvEDR1:1 MKIPFVTKWSHRSSEPAGPSNSAAAQQQQ PPPLSPSAPSRSPPVASTEAAGDEFILQEEEYQMQLALALSASASGAEGAGDPDGEQIRKA 90
TaEDR1:87 KLMSLGKGHPVTNSDRGGGDTPESLSRRYRDYNFLDYNEKVIDGFYDVFGLSAGSSGQGKIPSLAELQMSIGDLGYEVIVVDYKFDNALQ 176
KLMSLGKG?PVTNSD?GGG?T?ESLSRRYRDYNFLDYNEKVIDGFYD+FG?SA?SSG?GKIPSLAEL?MSIGDLGYEVIVVDYKFDNALQ
HvEDR1:91 KLMSLGKG DPVTNSDLGGGYTAESLSRRYRDYNFLDYNEKVIDGFYD IFGPSAESSGHGKIPSLAELHMSIGDLGYEVIVVDYKFDNALQ 180
TaEDR1:177 EMKEVAECCLLGCPDITVLVRRIAEVVADHMGGPVIDANEMITRWLSKSIEQRTSHQTSLLHIGSIEIGLSRHRALLFKILADIVGIPCK 266
EMKEVAECCLLGCPDITVLVRRIAEVVADGNGGPVIDANEMITRWLSKSIEQRTSHQTSLLHIGSIEIGLSRHRALLFKILAD+VGIPCK
HvEDR1:181 EMKEVAECCLLGCPDITVLVRRIAEVVADHMGGPVIDANEMITRWLSKSIEQRTSHQTSLLHIGSIEIGLSRHRALLFKILAD MVGIPCK 270
TaEDR1:267 LVKGSHYTGVEDDAINIIKMDDKREFLVDVMAAPGTLIPADVFNSKGTPFNFSQTLGQNQVVESASNIEDDPVALQSEHKRNQGHMFANN 356
LVKGSHYTGV?DDAINIIKMD+KREFLVDVMAAPGTLIPADVFNSKGTPFNFSQTLGQNQVVESASNIEDDPVALQSEH+ QGHMFANN
HvEDR1:271 LVKGSHYTGV VDDAINIIKMD NKREFLVDVMAAPGTLIPADVFNSKGTPFNFSQTLGQNQVVESASNIEDDPVALQSEH EHYQGHMFANN?360
TaEDR1:357 NRISVNLSSYENTMTAGSSASEPGTLDPRMQLGKTSTLPSAPSKQKKNLQLITDSHETEESRKLFVELDPFNAIESGKSSLAFKGLNNRN 446
+R+S?NLSSYENTMTAGSSASEPGTL GK?STL APSKQKKNLQLI?DSHE+ESR LF?E?DPFNA?ESGKSSLAFKGLNNRN
HvEDR1:361? DRVSDNLSSYENTMTAGSSASEPGTL ------GKASTLAGAPSKQKKNLQLIPDSHEIDESRNLFAEFDPFNATESGKSSLAFKGLNNRN 444
TaEDR1:447 NEFQRRRENVVPPSVRSQQPLVMKNWSACNDISNNKQYNVADGSVPRRNATDNASSSQLALSTAKHYNSNVRELNDRVYAAPARNYDNKI 536
++F+RRRENVVPPS?RSQQPLV?KNWSACNDISNNKQYNVADGSVPRRNATDNASSSQLALSTAKHYN?NVRELNDR+YAAPARNYDN+I
HvEDR1:445
Figure C20051001780600101
TaEDR1:537 VGTSAMAKALTGECPDRSQVPPGLYYDKMLGTSSMNAASTSGIGKVAEKDPHNDPGKGPIYSRFDGELSKNAQGFTPERDEHKENCGSHD 626
+GTSAMAKA?TG+C?DRSQVPPGLYYDKMLGTSSMN?AS+SGIGKVAEKD ND KGPIYSRFDGELSKNAQGFTPERDEHKENCGS+D
HvEDR1:535? IGTSAMAKASTGDCLDRSQVPPGLYYDKMLGTSSMN TASSSGIGKVAEKDLQNDLEKGPIYSRFDGELSKNAQGFTPERDEHKENCGS YD?624
TaEDR1:627 HKMLYPDPRKSPLDRFMDRPRQSIECVFPSQVGSNKADMVLDEVSECEILWEDLVIDERIGIGSYGEVYHADWNGTEVAVKKFLDQEFYG 716
H+ML+PDPRKSPLDRFMDRPRQ+IECV?PSQVGS+K?D+VLDEVSECEILWEDLVIDERIGIGSYGEVYHADWNGTEVAVKKFLDQEFYG
HvEDR1:625? HRMLHPDPRKSPLDRFMDRPRQ NIECVSPSQVGSSKVDLVLDEVSECEILWEDLVIDERIGIGSYGEVYHADWNGTEVAVKKFLDQEFYG 714
TaEDR1:717 DALEEFRCEVRIMRRLRHPNIVLFMGAVTRPPHLSIVSEYLPRGSLYKIIHRPNCQIDEKRRIKMALDVARGMNCLHTSVPTIVHRDLKS 806
DALEEFRCEVRIMRRLRHPNIVLFMGAVTRPPHLSIVSEYLPRGSLYKIIHRPNCQIDEKRRIKMALDVARGMNCLHTSVPTIVHRDLKS
HvEDR1:715 DALEEFRCEVRIMRRLRHPNIVLFMGAVTRPPHLSIVSEYLPRGSLYKIIHRPNCQIDEKRRIKMALDVARGMNCLHTSVPTIVHRDLKS 804
TaEDR1:807 PNLLVDDNWTVKVCDFGLSRLKHSTFLSSKSTAGTPEWMAPEVLRNEQSNEKCDIYSFGVILWELATLRKPWHGMNQMQVVGAVGFQDRR 896
PNLLVDDNWTVKVCDFGLSRLKHSTFLSSKSTAGTPEWMAPEVLRNEQSNEKCDIYSFGVILWELATLRKPWHGMNQMQVVGAVGFQDRR
HvEDR1:805 PNLLVDDNWTVKVCDFGLSRLKHSTFLSSKSTAGTPEWMAPEVLRNEQSNEKCDIYSFGVILWELATLRKPWHGMNQMQVVGAVGFQDRR 894
TaEDR1:897 LDIPKEVDPIVASIIRDCWQKDPNLRPSFIQLTSYLKTLQRLVIPSHQETASNHVPYEISLYR?959
LDIPKEVDPIVASIIRDCWQKDPNLRPSFIQLTSYLKTLQRLVIPSHQETASNHVPYEISLYR
HvEDR1:895 LDIPKEVDPIVASIIRDCWQKDPNLRPSFIQLTSYLKTLQRLVIPSHQETASNHVPYEISLYR?957
Wheat TaEDR1 albumen and the proteic comparison of barley HvEDR1, discrepant part marks with underscore, Represent a uncertain amino-acid residue, from comparative result as can be seen: the approach of wheat antidisense related gene TaEDR 1 of the present invention is one of important channel of disease-resistant wheat.
Embodiment two: the application of wheat antidisense related gene TaEDR 1 of the present invention on the transgenosis breeding for disease resistance.
TaEDR1 gene enzyme from the pGEM-T easy carrier is scaled off, or increase with high-fidelity Taq-DNA polysaccharase and gene-specific primer, TaRDR1 is connected the downstream of corn ubiquitin efficient promoter Ubi (Ubiquitin-1) with the T4-DNA ligase enzyme, be inserted into again in the pCAMBIA-Bar plant expression vector, thereby be built into pCAMBIA-Ubi-TaEDR1 expression vector plasmid.This expression vector is transformed in the intestinal bacteria DH10B bacterial strain breeds, extract the plasmid of this carrier, can carry out the transgenosis breeding for disease resistance.(the high pressure ammonia particle gun as Bole company PDS-1000/He) is transformed into high yield, high-quality with pCAMBIA-Ubi-TaEDR1 expression vector plasmid, but in the rataria callus of the wheat breed of disease resistance difference by particle gun.Utilize the selection markers gene on the expression vector---anti-herbicide gene Bar, in being added with the MS solid medium of weedicide PPT, screen, obtain the transgenic calli of anti-PPT.To screen the division culture medium (PPT that contains 3mg/L that the transgenic calli that obtains forwards new induced tissue differentiation to again; The Zeatin of 1mg/L, IAA with 1mg/L, 1/2 MS solid medium) cultivates in, induce and produce new regeneration plant, so just obtained transgenic wheat, detected by the offspring of round pcr, selected gene pure transfer-gen plant with gene-specific primer, the desirable transgenic strain that disease resistance is good can carry out field planting, popularization.
Embodiment three: the application of wheat antidisense related gene TaEDR 1 on the synthetic new disease-resistant related gene of design.
Modern DNA synthetic technology can be synthesized full-length gene, therefore, difference according to TaEDR1 gene and HvEDR1 gene, the synthetic back of cDNA before the HvEDR1 genes encoding 45 amino acids residues is connected to the gene cDNA terminal portions with TaEDR1 genes encoding 41 amino acids residues, has so just synthesized brand-new gene.Then, whether the encoder block (ORF) by the sequence verification synthetic gene is correct.Gene by checking can further be building up in the plant expression vector, detect by the disease resistance of transgenic technology synthetic gene, synthetic gene with good resistance characteristic of disease can be used for the cultivation of transgenosis anti-disease wheat new variety, and the structure of its plant expression vector is described identical with embodiment two with genetically modified concrete operations.
Embodiment four: the application of wheat antidisense related gene TaEDR 1 on gene chip is made.
With the TaEDR1 gene cDNA clone is template, obtain the DNA product with the M13 universal primer on gene-specific primer or the pGEM-T easy carrier by pcr amplification, product is carried out purifying, after the dissolving, make the wheat cdna chip according to the gene chip production process, be used for the disease-resistant correlative study of Denging, as can put automatically with the BioRobotics of Britain the film instrument with specimens point on 8 * 12 centimetres nylon membrane, 2 points of each sample spot, the DNA amount of each point is several nanograms, with house-keeping gene on the time point (as 18sRNA and Actin gene) as the stably express genetic contrast, the mRNA of experimental study material extract the back with isotropic substance (as 33P) mark is hybridized with gene chip as probe, analyzes hybridization signal with registering instrument, analyzes TaEDR1 expression of gene variation under the different experimental conditions, the relation that research TaEDR1 gene and disease-resistant etc. reacts according to the power of hybridization signal.
Embodiment five: the expression analysis method of wheat antidisense related gene TaEDR 1 in the mildew-resistance reaction:
The concrete grammar of expression analysis: (1) stably express crt gene---the amplification of wheat actin gene (Actin): according to sequence (the GenBank query ID: gi:48927617) designed a pair of primer of the Actin gene cDNA fragment among the GenBank, WAC-F:5 '-GTTCCAATCTATGAGGGATACACGC-3 ' and WAC-R:5 '-GAACCTCCACTGAGAACAACATTACC-3 ', reverse transcription PCR carries out on Perkin-Elmer 9600 gene-amplificative instraments, reaction solution is 25 μ L systems, the quantitative equivalent cDNA of process that comprises 3 μ L, 0.4 every kind of gene-specific primer of μ mol/L, the dNTP of 200 μ mol/L, 1 times PCR damping fluid, the MgCl of 1.5mmol/L 2, the Taq-DNA polysaccharase of 1-2 unit; Amplification program is: 94 ℃ of sex change 3 minutes; 94 ℃ of sex change of 18 round-robin 15 seconds were annealed 15 seconds for 55 ℃, and 72 ℃ were extended 1 minute; Extended 7 minutes at 72 ℃ at last.Amplified production is electrophoretic separation in 1% sepharose, the dyeing of yttrium bromide ingot, and ultraviolet lamp is observed, is taken a picture down, and the specific amplification products of wheat actin gene is that 422bp is long.(2) amplification of wheat TaEDR1 gene: method is identical with aforesaid method, and just the annealing temperature difference of amplification is 50 ℃, and gene-specific primer is: p1:
5 '-TTTCGTTGTGAAGTGAGG-3 '; P2:5 '-ATGGCTTTCTTAGTGTTGC-3 ', the specific amplification products of TaEDR1 gene are that 469bp is long.
Sequence table
<110〉Agricultural University Of He'nan
<120〉wheat antidisense related gene TaEDR 1 and application thereof
<160>2
<210>1
<211>3050
<212>DNA
<213〉common wheat (Triticum aestivum L.)
<400>1
cgccaaactg?aaataaccaa?gagaggaaat?attttcgaaa?cccaaaccct?aggtgctcgt 60
ggggagcgcc?atgaagatcc?cgtttgtgac?caagtggtcg?caccgatcca?gcgagcccgc 120
ggggccgtcg?aattcggctg?cagcgcagca?gcagcagcag?cagcaggcgc?cgtctcctcc 180
tcctcccgtg?gcgtcgacag?aggcggcagg?ggatgagttc?attctgcagg?aggaagagta 240
ccagatgcaa?ctggcgttgg?cgctatcagc?gtcggcgtcg?ggcgccgagg?gcgctgggga 300
tcccgacggg?gagcagatca?gaaaggcgaa?gctgatgagc?ctcgggaagg?gccacccggt 360
caccaacagc?gatcgtggcg?ggggagacac?cccggagtcg?ctctcccgcc?gttacaggga 420
ctataacttt?cttgattaca?atgagaaagt?aattgatgga?ttctacgacg?tatttggcct 480
ctctgcggga?tcatctgggc?agggcaaaat?accttcactg?gcagagcttc?agatgagcat 540
tggggatctt?ggatatgaag?taattgtggt?tgactataaa?tttgataatg?ctctgcagga 600
gatgaaggaa?gtagcagaat?gctgcctgtt?gggctgtcct?gacattacag?tattggtgcg 660
acgaatagct?gaagttgttg?cagatcacat?gggtggtcca?gtgatcgatg?caaatgaaat 720
gatcactagg?tggttgagca?aaagcattga?gcagaggaca?tcacaccaga?caagccttct 780
gcatattggc?agtatagaga?taggcttgtc?tcgccatcgt?gccttacttt?tcaagattct 840
tgctgatatt?gttggtatcc?cttgcaagct?ggttaaaggg?agtcattaca?ctggtgttga 900
agatgacgct?attaacataa?taaagatgga?tgacaaaagg?gagtttttgg?tggatgttat 960
ggctgctcca?gggactctca?ttccagcaga?tgtctttaat?tcaaagggta?ctccattcaa 1020
cttcagtcaa?acattgggtc?agaatcaggt?ggtggagtca?gcaagtaaca?tcgaagatga 1080
cccagttgca?ttacagtcag?agcataaacg?taaccaaggg?catatgtttg?ccaataataa 1140
tcggatctca?gtcaatctat?caagctatga?gaatacaatg?accgctggaa?gtagtgctag 1200
tgaacctggg?acattggacc?ctaggatgca?attaggtaaa?acatcaactt?tgcctagtgc 1260
tccttccaag?cagaagaaga?atctgcaatt?gattacagac?tctcatgaaa?ctgaagagtc 1320
ccgaaaacta?tttgtggagt?tagatccttt?caatgctatt?gaatctggga?aaagctcatt 1380
ggcattcaag?ggattaaata?atagaaacaa?tgaattccaa?aggcgtagag?agaatgtagt 1440
cccaccatct?gtaagatctc?aacagccatt?ggtgatgaaa?aactggtctg?cttgcaatga 1500
catttccaac?aacaagcaat?acaatgttgc?tgatgggtca?gttcctcgga?gaaatgccac 1560
tgacaatgca?tcgtcatctc?agttggcgtt?gtcaactgca?aagcattaca?attccaatgt 1620
tagagagcta?aacgatagag?tgtatgcagc?acctgctcgt?aattatgaca?acaagatagt 1680
tggtacctcg?gctatggcca?aagcattgac?tggagagtgc?cctgacagat?cacaggtgcc 1740
acctggtctt?tattatgaca?agatgcttgg?tacctcttct?atgaatgcag?cttctacatc 1800
cggaatcggg?aaagttgcag?aaaaggaccc?tcataatgat?ccgggaaaag?gtcccatcta 1860
ttctagattt?gatggtgaac?tttctaaaaa?tgctcaagga?tttactcccg?aaagggatga 1920
gcacaaggaa?aattgtggca?gtcatgacca?caaaatgtta?tatcctgatc?caagaaagtc 1980
ccctcttgac?agattcatgg?acaggccaag?gcagagcata?gaatgtgttt?ttccatccca 2040
agttggatca?aataaggctg?acatggtgtt?ggatgaagtg?tctgaatgtg?aaatcctttg 2100
ggaagatctt?gtaatcgatg?aaagaattgg?cataggttca?tatggagaag?tctaccatgc 2160
tgattggaat?ggaactgaag?tagctgtaaa?gaagttcttg?gatcaagagt?tctatggtga 2220
tgctttagag?gaatttcgtt?gtgaagtgag?gattatgcgt?cggctccgtc?atccaaatat 2280
tgttctcttt?atgggtgcag?taacacggcc?tccacactta?tctattgtat?cagaatatct 2340
tccaagggga?agcttatata?agatcattca?tcgccctaat?tgccaaatcg?acgagaagcg 2400
taggattaaa?atggcccttg?atgtggccag?aggcatgaat?tgtcttcata?ccagtgtacc 2460
aacaattgtt?caccgggatc?taaaatcacc?aaacttgctg?gttgacgata?attggactgt 2520
gaaggtctgt?gatttcggac?tttcacgtct?gaagcacagt?acatttttgt?catcaaaatc 2580
cactgccggg?actcctgagt?ggatggcacc?agaggttttg?cggaatgagc?aatccaatga 2640
gaagtgtgat?atttacagct?ttggtgttat?cctgtgggag?ctagcaacac?taagaaagcc 2700
atggcatggg?atgaaccaaa?tgcaagttgt?gggcgcagtt?ggcttccagg?accgacggct 2760
tgacattcca?aaagaagtag?atcctatagt?tgcatcaatt?atacgtgatt?gctggcagaa 2820
ggatccaaac?ttgcgtcctt?ctttcatcca?attaactagc?tacctgaaga?cattgcaaag 2880
gcttgtaatc?ccttcacatc?aggagacagc?gagcaaccat?gtaccctatg?aaatatcttt 2940
atatcggtga?accgcacatc?tctaccccgg?ggttgtcacc?caccaagtgt?gaataagcag 3000
taattatgat?tttgtcatcg?aaaaaaaaaa?aaaaaaaaaa?aaaaaaaaaa 3050
<210>2
<211>959
<212>PRT
<213〉common wheat (Triticum aestivum L.)
<400>2
Met?Lys?Ile?Pro?Phe?Val?Thr?Lys?Trp?Ser?His?Arg?Ser?Ser?Glu
1 5 10 15
Pro?Ala?Gly?Pro?Ser?Asn?Ser?Ala?Ala?Ala?Gln?Gln?Gln?Gln?Gln
20 25 30
Gln?Gln?Ala?Pro?Ser?Pro?Pro?Pro?Pro?Val?Ala?Ser?Thr?Glu?Ala
35 40 45
Ala?Gly?Asp?Glu?Phe?Ile?Leu?Gln?Glu?Glu?Glu?Tyr?Gln?Met?Gln
50 55 60
Leu?Ala?Leu?Ala?Leu?Ser?Ala?Ser?Ala?Ser?Gly?Ala?Glu?Gly?Ala
65 70 75
Gly?Asp?Pro?Asp?Gly?Glu?Gln?Ile?Arg?Lys?Ala?Lys?Leu?Met?Ser
80 85 90
Leu?Gly?Lys?Gly?His?Pro?Val?Thr?Asn?Ser?Asp?Arg?Gly?Gly?Gly
95 100 105
Asp?Thr?Pro?Glu?Ser?Leu?Ser?Arg?Arg?Tyr?Arg?Asp?Tyr?Asn?Phe
110 115 120
Leu?Asp?Tyr?Asn?Glu?Lys?Val?Ile?Asp?Gly?Phe?Tyr?Asp?Val?Phe
125 130 135
Gly?Leu?Ser?Ala?Gly?Ser?Ser?Gly?Gln?Gly?Lys?Ile?Pro?Ser?Leu
140 145 150
Ala?Glu?Leu?Gln?Met?Ser?Ile?Gly?Asp?Leu?Gly?Tyr?Glu?Val?Ile
155 160 165
Val?Val?Asp?Tyr?Lys?Phe?Asp?Asn?Ala?Leu?Gln?Glu?Met?Lys?Glu
170 175 180
Val?Ala?Glu?Cys?Cys?Leu?Leu?Gly?Cys?Pro?Asp?Ile?Thr?Val?Leu
185 190 195
Val?Arg?Arg?Ile?Ala?Glu?Val?Val?Ala?Asp?His?Met?Gly?Gly?Pro
200 205 210
Val?Ile?Asp?Ala?Asn?Glu?Met?Ile?Thr?Arg?Trp?Leu?Ser?Lys?Ser
215 220 225
Ile?Glu?Gln?Arg?Thr?Ser?His?Gln?Thr?Ser?Leu?Leu?His?Ile?Gly
230 235 240
Ser?Ile?Glu?Ile?Gly?Leu?Ser?Arg?His?Arg?Ala?Leu?Leu?Phe?Lys
245 250 255
Ile?Leu?Ala?Asp?Ile?Val?Gly?Ile?Pro?Cys?Lys?Leu?Val?Lys?Gly
260 265 270
Ser?His?Tyr?Thr?Gly?Val?Glu?Asp?Asp?Ala?Ile?Asn?Ile?Ile?Lys
275 280 285
Met?Asp?Asp?Lys?Arg?Glu?Phe?Leu?Val?Asp?Val?Met?Ala?Ala?Pro
290 295 300
Gly?Thr?Leu?Ile?Pro?Ala?Asp?Val?Phe?Asn?Ser?Lys?Gly?Thr?Pro
305 310 315
Phe?Asn?Phe?Ser?Gln?Thr?Leu?Gly?Gln?Asn?Gln?Val?Val?Glu?Ser
320 325 330
Ala?Ser?Asn?Ile?Glu?Asp?Asp?Pro?Val?Ala?Leu?Gln?Ser?Glu?His
335 340 345
Lys?Arg?Asn?Gln?Gly?His?Met?Phe?Ala?Asn?Asn?Asn?Arg?Ile?Ser
350 355 360
Val?Asn?Leu?Ser?Ser?Tyr?Glu?Asn?Thr?Met?Thr?Ala?Gly?Ser?Ser
365 370 375
Ala?Ser?Glu?Pro?Gly?Thr?Leu?Asp?Pro?Arg?Met?Gln?Leu?Gly?Lys
380 385 390
Thr?Ser?Thr?Leu?Pro?Ser?Ala?Pro?Ser?Lys?Gln?Lys?Lys?Asn?Leu
395 400 405
Gln?Leu?Ile?Thr?Asp?Ser?His?Glu?Thr?Glu?Glu?Ser?Arg?Lys?Leu
410 415 420
Phe?Val?Glu?Leu?Asp?Pro?Phe?Asn?Ala?Ile?Glu?Ser?Gly?Lys?Ser
425 430 435
Ser?Leu?Ala?Phe?Lys?Gly?Leu?Asn?Asn?Arg?Asn?Asn?Glu?Phe?Gln
440 445 450
Arg?Arg?Arg?Glu?Asn?Val?Val?Pro?Pro?Ser?Val?Arg?Ser?Gln?Gln
455 460 465
Pro?Leu?Val?Met?Lys?Asn?Trp?Ser?Ala?Cys?Asn?Asp?Ile?Ser?Asn
470 475 480
Asn?Lys?Gln?Tyr?Asn?Val?Ala?Asp?Gly?Ser?Val?Pro?Arg?Arg?Asn
485 490 495
Ala?Thr?Asp?Asn?Ala?Ser?Ser?Ser?Gln?Leu?Ala?Leu?Ser?Thr?Ala
500 505 510
Lys?His?Tyr?Asn?Ser?Asn?Val?Arg?Glu?Leu?Asn?Asp?Arg?Val?Tyr
515 520 525
Ala?Ala?Pro?Ala?Arg?Asn?Tyr?Asp?Asn?Lys?Ile?Val?Gly?Thr?Ser
530 535 540
Ala?Met?Ala?Lys?Ala?Leu?Thr?Gly?Glu?Cys?Pro?Asp?Arg?Ser?Gln
545 550 555
Val?Pro?Pro?Gly?Leu?Tyr?Tyr?Asp?Lys?Met?Leu?Gly?Thr?Ser?Ser
560 565 570
Met?Asn?Ala?Ala?Ser?Thr?Ser?Gly?Ile?Gly?Lys?Val?Ala?Glu?Lys
575 580 585
Asp?Pro?His?Asn?Asp?Pro?Gly?Lys?Gly?Pro?Ile?Tyr?Ser?Arg?Phe
590 595 600
Asp?Gly?Glu?Leu?Ser?Lys?Asn?Ala?Gln?Gly?Phe?Thr?Pro?Glu?Arg
605 610 615
Asp?Glu?His?Lys?Glu?Asn?Cys?Gly?Ser?His?Asp?His?Lys?Met?Leu
620 625 630
Tyr?Pro?Asp?Pro?Arg?Lys?Ser?Pro?Leu?Asp?Arg?Phe?Met?Asp?Arg
635 640 645
Pro?Arg?Gln?Ser?Ile?Glu?Cys?Val?Phe?Pro?Ser?Gln?Val?Gly?Ser
650 655 660
Asn?Lys?Ala?Asp?Met?Val?Leu?Asp?Glu?Val?Ser?Glu?Cys?Glu?Ile
665 670 675
Leu?Trp?Glu?Asp?Leu?Val?Ile?Asp?Glu?Arg?Ile?Gly?Ile?Gly?Ser
680 685 690
Tyr?Gly?Glu?Val?Tyr?His?Ala?Asp?Trp?Asn?Gly?Thr?Glu?Val?Ala
695 700 705
Val?Lys?Lys?Phe?Leu?Asp?Gln?Glu?Phe?Tyr?Gly?Asp?Ala?Leu?Glu
710 715 720
Glu?Phe?Arg?Cys?Glu?Val?Arg?Ile?Met?Arg?Arg?Leu?Arg?His?Pro
725 730 735
Asn?Ile?Val?Leu?Phe?Met?Gly?Ala?Val?Thr?Arg?Pro?Pro?His?Leu
740 745 750
Ser?Ile?Val?Ser?Glu?Tyr?Leu?Pro?Arg?Gly?Ser?Leu?Tyr?Lys?Ile
755 760 765
Ile?His?Arg?Pro?Asn?Cys?Gln?Ile?Asp?Glu?Lys?Arg?Arg?Ile?Lys
770 775 780
Met?Ala?Leu?Asp?Val?Ala?Arg?Gly?Met?Asn?Cys?Leu?His?Thr?Ser
785 790 795
Val?Pro?Thr?Ile?Val?His?Arg?Asp?Leu?Lys?Ser?Pro?Asn?Leu?Leu
800 805 810
Val?Asp?Asp?Asn?Trp?Thr?Val?Lys?Val?Cys?Asp?Phe?Gly?Leu?Ser
815 820 825
Arg?Leu?Lys?His?Ser?Thr?Phe?Leu?Ser?Ser?Lys?Ser?Thr?Ala?Gly
830 835 840
Thr?Pro?Glu?Trp?Met?Ala?Pro?Glu?Val?Leu?Arg?Asn?Glu?Gln?Ser
845 850 855
Asn?Glu?Lys?Cys?Asp?Ile?Tyr?Ser?Phe?Gly?Val?Ile?Leu?Trp?Glu
860 865 870
Leu?Ala?Thr?Leu?Arg?Lys?Pro?Trp?His?Gly?Met?Asn?Gln?Met?Gln
875 880 885
Val?Val?Gly?Ala?Val?Gly?Phe?Gln?Asp?Arg?Arg?Leu?Asp?Ile?Pro
890 895 900
Lys?Glu?Val?Asp?Pro?Ile?Val?Ala?Ser?Ile?Ile?Arg?Asp?Cys?Trp
905 910 915
Gln?Lys?Asp?Pro?Asn?Leu?Arg?Pro?Ser?Phe?Ile?Gln?Leu?Thr?Ser
920 925 930
Tyr?Leu?Lys?Thr?Leu?Gln?Arg?Leu?Val?Ile?Pro?Ser?His?Gln?Glu
935 940 945
Thr?Ala?Ser?Asn?His?Val?Pro?Tyr?Glu?Ile?Ser?Leu?Tyr?Arg
950 955 959

Claims (5)

1. wheat antidisense related gene TaEDR 1, its base sequence is shown in SEQ ID NO:1.
2. coded albumen of the described wheat antidisense related gene TaEDR 1 of claim 1, its aminoacid sequence is shown in SEQ ID NO:2.
3. the application of wheat antidisense related gene TaEDR 1 in the plant disease-resistant improvement.
4. the application of wheat antidisense related gene TaEDR 1 in synthetic new disease-resistant related gene.
5. the application of wheat antidisense related gene TaEDR 1 in the wheat cdna chip manufacturing.
CNB2005100178062A 2005-07-20 2005-07-20 Wheat antidisense related gene TaEDR1 and its application Expired - Fee Related CN1295334C (en)

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