CN1216908C - Arabidopsis thaliana BVD1 gene and its use - Google Patents
Arabidopsis thaliana BVD1 gene and its use Download PDFInfo
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Abstract
The present invention relates to a gene cloned from changed mutant, namely bushy and dwarf 1(bud1) of arabidopsis strains. Through identification, the gene which is named as BUD1 influences the metabolism of indoleacetic acid (IAA) of plant strains so as to further control the forms of the strains. Transgenic experiments indicate that excessive expression of the BUD1 can control the form of plant strains. The comparison analysis of amino acid sequences indicates that the gene is unknown MAPKK-AtMKK7 in an MAPK cascade system. The high expression of the BUD1 in arabidopsis causes dwarf forms and the lose of apical dominance, which indicates that the strain forms of plants can be changed by a genetic engineering technology. The gene has important application value in aspects of adjusting mechanism of the metabolism of growth hormone and purposefully adjusting the characteristic of strain forms of plants.
Description
Technical field
The invention belongs to plant genetic engineering field.Specifically, the present invention relates to a kind of BUD1 (BUSHY AND DWARF 1) gene that activates the labeling acts clone, and utilize transgenic experiments to identify this gene; Also relate to and contain this gene and its and have homologous gene, the related vector of the homologous sequence of similar functions and other species and utilize this gene or characters with plant such as the apical dominance of its functional analogue regulation and control plant, plant height.
Technical background
The plant type of plant is decided by the branching system of stem to a great extent.The structure of whole stem determines jointly by resulting from other meristematic tissue that produces brephic stem apical meristem and postembryonic period.The apical meristem of stem has determined the main shaft of plant, derives from the configuration that other merismatic side shoots have further been modified plant again.The branching process of stem generally comprises two etap: the formation of axillary meristem and the growth of axillalry bud in the axil.Shimizu-Sato S thinks that in calendar year 2001 in many species, the growth of axillalry bud is suppressed by stem or main rachis, i.e. " apical dominance ".Apical dominance is an important economical character.On gardening, flowers and trees often need pinching, and the terminal bud of cutting branch promotes axillary bud growth to remove apical dominance, forms the plentiful strain clump of multi-branched, and plant is controlled at the ideal height, improves ornamental value.
Apical dominance is a process that is subjected to phytohormone Regulation.The research of past to apical dominance mainly is to utilize physiological method, applies plant hormone and hormone-content by external source and measures and show plant hormone, as growth hormone and phytokinin, plays an important role in the control to this process.Klee H thinks that in 1991 growth hormone has the effect of inhibition to the growth of axillalry bud, and phytokinin then promotes axillary bud growth.Whether axillary bud growth is decided by the ratio of these two kinds of hormones rather than any absolute content.But traditional Physiologic Studies method can not accurately be controlled absorbed dose and the position of plant to exogenous hormone.By contrast, the mutant of separation branching pattern change, evaluation mutator gene function just become the more strong instrument of research apical dominance mechanism.The gene that influences apical dominance of having cloned at present seldom, comprise Leyser H in clone's in 1993 Arabidopis thaliana growth hormone signal transduction path the AXR1 gene and Tantikanjana T the calendar year 2001 clone with the synthetic relevant SUPERSHOOT gene of phytokinin.The present invention passes through to activate the gene BUD1 of controlled Arabidopis thaliana apical dominance of labeling acts and plant height, and has identified the function of this gene by transgenic experiments.
Summary of the invention
At above-mentioned research background, the purpose of this invention is to provide a kind of new gene of from arabidopsis mutant body bud1, cloning, BUD1, the dna sequence dna shown in seq ID No1 also comprises the gene order that has 70% homology with the dna sequence dna shown in the seq IDNo.1 at least; Be also included within the mutant allele or the derivative that add, replace, insert or delete one or more Nucleotide and produce, also contain the gene order that has identical function and can reach the object of the invention.A kind of protein shown in seq ID No2 also is provided among the present invention, wherein carries out one or several amino acid whose replacement, insertion or the disappearance functional analogue that amino acid obtained.The present invention also comprises the functional analogue that has the homology that (comprises 70%) more than 70% with the aminoacid sequence shown in the seq ID No2.
Another object of the present invention provides a kind of method of carrying out the high-efficiency plant genetic transformation with BUD1.Any can expression by the polypeptide of above-mentioned nucleic acid sequence encoding or the carrier of homology analogue.The present invention also provides a kind of host cell that contains above expression vector.Host cell comprises intestinal bacteria, Agrobacterium and vegetable cell.
Another object of the present invention provides a kind of the utilization and activates the labeling acts conversion to change the method for plant forms.
Realize that concrete technological step of the present invention is as follows:
One Arabidopis thaliana plant type changes separation and the genetic analysis of mutant bud1
It is to utilize justice/antisense expression (SARE) system [7] to transform the environmental plant of wild-type Arabidopis thaliana Col-0, the mutant bud1 that the phenotype that obtains by a large amount of screenings is stable again that the used Arabidopis thaliana plant type of the present invention changes mutant.By to bud1 and wild-type Arabidopis thaliana Col-0 hybridization F1 and the F2 analysis for phenotypic segregation ratio, we determine that bud1 is a semidominance mutant that meets single-gene control of heredity rule.The forfeiture of this mutant apical dominance, short and small, leaf roll is bent, heterozygote half is sterile, but makes solid amount increase, homozygote sterile fully because branch, as shown in Figure 1.The separation bud1 phenotype of gene BUD1 of two control plant forms and T-DNA be divided into from
For the separate targets gene, the present invention has chosen bud1 and wild-type Arabidopis thaliana Col-0 filial generation F2 10 wild-type plant and 70 mutant plant in generation, with EcoR I complete degestion genome, with the special 2 * 35S enhanser probe of T-DNA do Southern hybridization detect mutant phenotype and T-DNA be divided into from.The result shows, the T-DNA that contains 3 copies in the genetically modified mutant altogether at least inserts, wherein the band of 7kb and mutant be divided into from, as shown in Figure 2.The separation of T-DNA flanking sequence
Recovery is built into phage subgene group library through the DNA of the bud1 genome 6kb-8kb of EcoR I complete degestion section.To be transformed into plasmid, order-checking with the purpose clone that the special 2 * 35S enhanser probe screening of T-DNA obtains.According between the nt61451 and nt61452 of the sequencing result search Arabidopis thaliana database discovery T-DNA insertion Arabidopis thaliana first karyomit(e) BAC15H18 (Fig. 3).Insertion causes the overexpression (Fig. 4) of T-DNA one side one prediction MAPKK-AtMKK7.
The evaluation of BUD1 gene and functional analysis
Carry out transgenic experiments by the vacuum filtration method that Bechtold set up in 1993, the result shows that the present invention has obtained to simulate the transgenic arabidopsis of bud1 phenotype, proof the present invention has correctly cloned the BUD1 gene, and the overexpression of proof BUD1 gene can cause the change of plant type.Gene DNA sequence and aminoacid sequence are seen Seq ID No.1 and No.2.
Control to the plant plant type is to depend on artificial pruning substantially at present, if can realize by biotechnology will be more economical effective to the control of plant apical dominance, when particularly being applied to ornamental plant, because ornamental plant only supplies to view and admire but not is edible, with regard to its security, easilier go through to put on market and realize commercialization.The discovery of BUD1 gene makes the plant type of using gene engineering technique regulation and control plant become possibility with the clone.
Description of drawings
Be described in further detail understanding the present invention below in conjunction with accompanying drawing, but be not that the present invention is limited.
Fig. 1. the Arabidopis thaliana plant type changes the phenotype of mutant bud1. a left side: wild-type Arabidopis thaliana; In: the bud1 heterozygote; Right: the bud1 homozygote
Fig. 2 .bud1 phenotype and T-DNA are divided into from .A: wild-type, bud1 heterozygote and homozygote genomic dna are cut through EcoR I enzyme, 2 * 35S probe hybridization, show a bud1 and a 7kb band be divided into from; B: the strain that only contains the 7kb band is that genomic dna is cut through plurality of enzymes, and 2 * 35S probe hybridization shows 2 hybrid belts.
Fig. 3 .T-DNA inserts the site synoptic diagram
The overexpression .A:Nothern result of BUD1 gene among Fig. 4 bud1; B:RT-PCR result.
The dna sequence dna and the aminoacid sequence of Fig. 5 .BUD1 gene
Embodiment
Case study on implementation 1
1. arabidopsis mutant body bushy and dwarf 1 (bud1), original wild-type material is the Col-0 ecotype.
2. plant culturing
Arabidopis thaliana is cultivated and is adopted vermiculite to water with 1/3 B5 nutrient solution, and growth conditions is 23 ℃ of continuous illuminations, and intensity is 80-120 μ molm
-2Sec
-1The Arabidopis thaliana sterile culture adopts surface sterilization, and 70% alcohol surface disinfection soaked sterilization 10-15 minute, aseptic washing 3-4 time with 10%Bleach after 3-5 minute.Handle for 4 ℃ and grew placing on the 1/2 MS substratum to cultivate under the identical condition with program request behind the stratification in 4 days with vermiculite.
3.bud1 with being divided into of T-DNA from experiment
Hybridize F2 for choosing 10 wild-type plant and 70 mutant plant the colony from bud1 homozygote and wild-type Arabidopis thaliana, extract genome DNA.The method of the CTAB that Lijiayang nineteen ninety-five creates is adopted in the extraction of plant genome DNA.With genomic dna with restriction enzyme EcoR I complete degestion after in 1% sepharose electrophoresis, changeing film instrument (Biorad) with vacuum goes to it on nylon membrane, with the special 2 * 35S enhanser probe of T-DNA in 65 ℃ of hybridization 16-20 hour, after preciseness is washed film, scan and analyze with Phosphoimager.The result shows that the T-DNA that contains 3 copies in the genetically modified mutant altogether at least inserts, and wherein the band of 7kb and mutant are divided into from (Fig. 2).The mutant strain system of only containing this 7kb band is retained, and all research work all only utilize this strain system.This strain is that genomic dna is used other restriction enzymes again: when BamH I, Xho I, Sac I complete degestion, with 2 * 35S probe, two bands (Fig. 2) of mixing out, this two band does not separate in mutant, illustrates that this is two closely linked T-DNA fragments.Bud1 is the mutant that utilizes the SARE system to obtain, the SARE system imports the cDNA segment [7] of external source to transfer-gen plant, but reason owing to probability, sometimes be not connected with such cDNA on the carrier in conversion library, the sudden change that conversion causes may only be to insert sudden change, disturbs native gene to express with external source cDNA and has nothing to do.The Mutagen that the method for PCR is utilized to identify bud1 because of.With the transgenic plant genome is template, 5 '-ACCACGTCTTCAAAGCAAGTG-3 ' (from 2 * 35S enhanser) and 5 '-TATGATAATCATCGCAAGACCG-3 ' (from the NOS terminator) will obtain 2 PCR product bands for primer, because from the primer of 2 * 35S enhanser 2 binding sites are arranged on 2 * 35S enhanser.If the T-DNA segment is not with external source cDNA, product length is 0.2kb and 0.5kb.The product that obtains from bud1 is 0.2kb and 0.5kb, and the sequencing result proof does not wherein contain external source cDNA segment really.Therefore bud1 is an insertion mutant.The 7kb band that this sudden change and EcoR I enzyme are cut be divided into from.
4.T-DNA the separation of flanking sequence
From the DNA of sepharose recovery through the bud1 genome 6kb-8kb of EcoR I complete degestion section, be connected into λ ZAP II carrier (Stratagene product), be built into a phage subgene group library.Obtain 20 mono-clonals with 2 * 35S probe through the three-wheel screening.Each mono-clonal is changed into plasmid, the upgrading grain, enzyme is cut detection.The result shows that 20 mono-clonals demonstrate 2 kinds of enzymes and cut finger printing, represents 2 kinds of different plasmids.This is consistent with aforementioned Southern detected result.Get 2 kinds of plasmid 13-2-5 and 5-1-3 order-checking, search Arabidopis thaliana database finds that 13-2-5 contains the T-DNA fragment, insert the Arabidopis thaliana first karyomit(e) BAC15H18, left margin next-door neighbour nt61451 is at the MAPKK of a prediction (Atlg18350, AtMKK7) open reading frame (ORF) upstream 513bp; 5-1-3 also contains the T-DNA fragment, inserts the Arabidopis thaliana first karyomit(e) BAC15H18, and right margin next-door neighbour nt61452 is at the transcription factor 2.35kD of prediction subunit (Atlg18340) open reading frame upstream 1438bp.This shows between Arabidopis thaliana first karyomit(e) BAC15H18 nt61451 and nt61452 and inserted two T-DNA fragments.And sequencing result shows that these two T-DNA fragment sequences are identical, reverse repeated arrangement,
5.BUD1 expression amount detects
Total RNA extracts the acid guanidine thiocyanate-phenol-chloroform extraction method that adopts Wadsworth G to create in 1998.Get the 1g vegetable material and grind in liquid nitrogen, through guanidine thiocyanate, acid phenol and chloroform extracting, behind the ethanol sedimentation, with LiCL washing once, total RNA is dissolved in the water of nuclease free, measures behind the O.D. value constant volume in-20 ℃ of preservations.Utilize the method for RT-PCR, design gene-specific primer 5 '-GGATCCTCTCTCTTCTATTTCCATGGC-3 ', 5 '-GAGCTCACAAGCAGTCGGATCTAAAG-3 ' and 5 '-AGGAGATGTCTTCCGCTGATG-3 ', 5 '-AAGTCTTCATCGGATTCGTGTG-3 ', with the total RNA reverse transcription product of wild-type is increase respectively Atlg18350 and Atlg18340 cDNA fragment of template, and amplified production proves this gene itself through order-checking.Atlg18350 cDNA sequence is consistent with the prediction of arabidopsis gene group database.RT-PCR adopts SUPERS CRIPT first Strand Synthesis System (Gibco BRL product), and reaction system is 20 μ l.Wherein the total RNA of 2 μ g removes DNA impurity through the DNaseI degraded in advance, and 2 μ l reverse transcription products are used for pcr amplification subsequently, and amplification system is 50 μ l.The PCR reaction conditions is 94 ℃ of 3min., 35 circulations (94 ℃ of 1min., 58 ℃ of 1min., 72 ℃ of 1min.), 72 ℃ of 10min.Make probe with above-mentioned RT-PCR product and carry out Northern hybridization.With the total RNA of 10-15 μ g in denaturing formaldehyde glue behind the electrophoresis, be transferred on the nylon membrane, in vacuum oven in 80 ℃ roasting 2 hours with set, the Church buffer system that Northern hybridization adopts Church GM to create in 1984, and scan and analyze with Phosphoimager.The Northern results of hybridization shows that Atlg18350 is overexpression in bud1, then can not detect signal (Fig. 4) in wild-type.No matter and Atlg18340 equal amixia signal in bud1 or wild-type.We further verify Northern result (Fig. 4) by the RT-PCR method again, contrast the gene into ACTIN7.The used primer of amplification ACTIN7 gene is 5 '-TGGAATGGTGAAGGCTGGTTT-3 ' and 5 '-CTGTTGGAAGGTGCTGAGGGA-3 '.The expression pattern of Atlg18350 is with Northern unanimity as a result.The expression amount of Atlg18340 is not seen difference in mutant and wild-type.It is to activate the mutant that labeling acts causes that above data can support such conclusion: bud1 actual, and (Atlg18350 AtMKK7) is inserted into tag activation to the MAPKK of a prediction, and expression amount improves near the insertion site.We are this MAPKK called after BUD1.
6.BUD1 the acquisition of cDNA complete sequence
5 '/3 ' RACE kit (Roche product) has been used in the acquisition of 5 ' and 3 ' non-translational region of BUD1 gene.Used Auele Specific Primer is:
SP1(5’-GAGCTCAAGTATCATCACTCCG-3’)
SP2(5’-GCTAGTTGTTTCTCCGTAACGG-3’)
SP3(5’-CTGCTTCCTCTTCCGAGAAG-3’)
SP5(5’-CCGTTACGGAGAAACAACTAGC-3’)。
Case study on implementation 2,
Agrobacterium tumefaciens mediated arabidopsis thaliana transformation
The structure of the conversion plasmid of overexpression BUD1 is to be connected between the BamH I of pBI121 and the Sac I site by the genome PCR product with BUD1 to obtain, and the primer is 5 '-GGATCCTCTCTCTTCTATTTCCATGGC-3 ' and 5 '-GAGCTCACAAGCAGTCGGATAAAG-3 '.Change the conversion plasmid that makes up over to Agrobacterium GV3101 (pMP90) bacterial strain with electric shocking method, bacterium colony PCR identifies the vacuumizing method arabidopsis thaliana transformation Col-0 wild-type that the back is set up with Bechtold N1993, be sowed on the 1/2 MS substratum that contains kantlex (50ng/l) after the seed results, treat that T1 grows to during the time shift of 4-6 sheet lotus throne leaf buries for plant, get a lotus throne leaf in case of necessity and extract genomic dna and detect.After T1 gathered in the crops for individual plant, the part seed continued screening to observe the T2 separation case in generation with kantlex, and part is sowed in the vermiculite to obtain stable transgenic line.
bud1?patent.ST25
SEQUENCE?LISTING
＜110〉Chinese Academy of Sciences's heredity and developmental biology institute
＜120〉Arabidopis thaliana BUD1 gene and application thereof
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<223>
<400>1
accatcatca?ctctctctct?ctctcttcta?tttcc?atg?gct?ctt?gtt?cgt?aaa 53
Met?Ala?Leu?Val?Arg?Lys
1 5
cgc?cgt?caa?atc?aac?ctc?cgt?ctc?cct?gtc?cca?ccg?ctc?tct?gtt?cac 101
Arg?Arg?Gln?Ile?Asn?Leu?Arg?Leu?Pro?Val?Pro?Pro?Leu?Ser?Val?His
10 15 20
ctc?ccc?tgg?ttc?tcc?ttt?gcc?tca?tcc?acc?gcc?ccc?gtc?atc?aac?aac 149
Leu?Pro?Trp?Phe?Ser?Phe?Ala?Ser?Ser?Thr?Ala?Pro?Val?Ile?Asn?Asn
25 30 35
gga?atc?tca?gct?tcc?gat?gtc?gag?aaa?ctc?cac?gtt?ctc?gga?aga?gga 197
Gly?Ile?Ser?Ala?Ser?Asp?Val?Glu?Lys?Leu?His?Val?Leu?Gly?Arg?Gly
40 45 50
agc?agc?ggg?atc?gta?tac?aaa?gtc?cac?cac?aaa?acc?acg?ggg?gag?ata 245
Ser?Ser?Gly?Ile?Val?Tyr?Lys?Val?His?His?Lys?Thr?Thr?Gly?Glu?Ile
55 60 65 70
tac?gct?ctg?aaa?tca?gtc?aac?ggc?gac?atg?agt?cct?gct?ttc?aca?aga 293
Tyr?Ala?Leu?Lys?Ser?Val?Asn?Gly?Asp?Met?Ser?Pro?Ala?Phe?Thr?Arg
75 80 85
caa?tta?gcg?cgc?gag?atg?gag?atc?ctc?cgt?cgc?acg?gat?tct?cct?tat 341
Gln?Leu?Ala?Arg?Glu?Met?Glu?Ile?Leu?Arg?Arg?Thr?Asp?Ser?Pro?Tyr
90 95 100
bud1?patent.ST25
gtc?gtc?agg?tgt?caa?ggg?atc?ttc?gag?aaa?cca?atc?gtc?gga?gag?gtt 389
Val?Val?Arg?Cys?Gln?Gly?Ile?Phe?Glu?Lys?Pro?Ile?Val?Gly?Glu?Val
105 110 115
tcg?atc?ctc?atg?gag?tat?atg?gac?ggt?gga?aac?cta?gaa?tct?ctc?cgc 437
Ser?Ile?Leu?Met?Glu?Tyr?Met?Asp?Gly?Gly?Asn?Leu?Glu?Ser?Leu?Arg
120 125 130
ggc?gcc?gtt?acg?gag?aaa?caa?cta?gcg?gga?ttt?tcc?cgc?cag?att?ttg 485
Gly?Ala?Val?Thr?Glu?Lys?Gln?Leu?Ala?Gly?Phe?Ser?Arg?Gln?Ile?Leu
135 140 145 150
aaa?ggt?tta?agt?tat?ctc?cac?tca?ctc?aag?atc?gtt?cac?aga?gac?atc 533
Lys?Gly?Leu?Ser?Tyr?Leu?His?Ser?Leu?Lys?Ile?Val?His?Arg?Asp?Ile
155 160 165
aaa?cct?gcg?aat?cta?ctc?tta?aac?tcg?aga?aac?gaa?gtt?aaa?atc?gct 581
Lys?Pro?Ala?Asn?Leu?Leu?Leu?Asn?Ser?Arg?Asn?Glu?Val?Lys?Ile?Ala
170 175 180
gat?ttt?gga?gtg?agc?aaa?atc?att?acc?cga?tcg?tta?gat?tac?tgc?aat 629
Asp?Phe?Gly?Val?Ser?Lys?Ile?Ile?Thr?Arg?Ser?Leu?Asp?Tyr?Cys?Asn
185 190 195
tcc?tac?gtc?ggc?act?tgc?gct?tac?atg?agc?ccg?gag?aga?ttt?gac?tct 677
Ser?Tyr?Val?Gly?Thr?Cys?Ala?Tyr?Met?Ser?Pro?Glu?Arg?Phe?Asp?Ser
200 205 210
gcc?gcc?gga?gaa?aac?tcc?gat?gtt?tac?gca?ggc?gat?atc?tgg?agt?ttc 725
Ala?Ala?Gly?Glu?Asn?Ser?Asp?Val?Tyr?Ala?Gly?Asp?Ile?Trp?Ser?Phe
215 220 225 230
gga?gtg?atg?ata?ctt?gag?ctc?ttc?gtc?gga?cat?ttt?ccg?ttg?ctt?cct 773
Gly?Val?Met?Ile?Leu?Glu?Leu?Phe?Val?Gly?His?Phe?Pro?Leu?Leu?Pro
235 240 245
cag?gga?cag?aga?cct?gac?tgg?gcg?acg?tta?atg?tgc?gtg?gtg?tgc?ttt 821
Gln?Gly?Gln?Arg?Pro?Asp?Trp?Ala?Thr?Leu?Met?Cys?Val?Val?Cys?Phe
250 255 260
gga?gaa?cca?ccg?cgt?gcg?ccg?gaa?gga?tgt?tcc?gac?gag?ttt?agg?agt 869
Gly?Glu?Pro?Pro?Arg?Ala?Pro?Glu?Gly?Cys?Ser?Asp?Glu?Phe?Arg?Ser
265 270 275
ttt?gtt?gac?tgt?tgt?ctc?cgt?aaa?gaa?tcg?agt?gag?agg?tgg?acg?gcg 917
Phe?Val?Asp?Cys?Cys?Leu?Arg?Lys?Glu?Ser?Ser?Glu?Arg?Trp?Thr?Ala
280 285 290
tcg?cag?ctt?ctc?ggt?cac?cct?ttt?ctc?cgt?gaa?agt?ctt?tag
bud1?patent.ST25
Ser?Gln?Leu?Leu?Gly?His?Pro?Phe?Leu?Arg?Glu?Ser?Leu
295 300 305
atccgactgc?ttgtgatatt?acgggtctgg?atattttccg?ggtgacccaa?atcgcaatta 1019
ttatttttt?gtagaatttt?tgtatgatca?gaaatatgct?ctattgaaat?tcatctgacg 1079
att 1082
<210>2
<211>307
<212>PRT
<213>Atabidopsis?thaliana
<400>2
Met?Ala?Leu?Val?Arg?Lys?Arg?Arg?Gln?Ile?Asn?Leu?Arg?Leu?Pro?Val
1 5 10 15
Pro?Pro?Leu?Ser?Val?His?Leu?Pro?Trp?Phe?Ser?Phe?Ala?Ser?Ser?Thr
20 25 30
Ala?Pro?Val?Ile?Asn?Asn?Gly?Ile?Ser?Ala?Ser?Asp?Val?Glu?Lys?Leu
35 40 45
His?Val?Leu?Gly?Arg?Gly?Ser?Ser?Gly?Ile?Val?Tyr?Lys?Val?His?His
50 55 60
Lys?Thr?Thr?Gly?Glu?Ile?Tyr?Ala?Leu?Lys?Ser?Val?Asn?Gly?Asp?Met
65 70 75 80
Ser?Pro?Ala?Phe?Thr?Arg?Gln?Leu?Ala?Arg?Glu?Met?Glu?Ile?Leu?Arg
85 90 95
Arg?Thr?Asp?Ser?Pro?Tyr?Val?Val?Arg?Cys?Gln?Gly?Ile?Phe?Glu?Lys
100 105 110
Pro?Ile?Val?Gly?Glu?Val?Ser?Ile?Leu?Met?Glu?Tyr?Met?Asp?Gly?Gly
115 120 125
Asn?Leu?Glu?Ser?Leu?Arg?Gly?Ala?Val?Thr?Glu?Lys?Gln?Leu?Ala?Gly
bud1?patent.ST25
130 135 140
Phe?Ser?Arg?Gln?Ile?Leu?Lys?Gly?Leu?Ser?Tyr?Leu?His?Ser?Leu?Lys
145 150 155 160
Ile?Val?His?Arg?Asp?Ile?Lys?Pro?Ala?Asn?Leu?Leu?Leu?Asn?Ser?Arg
165 170 175
Asn?Glu?Val?Lys?Ile?Ala?Asp?Phe?Gly?Val?Ser?Lys?Ile?Ile?Thr?Arg
180 185 190
Ser?Leu?Asp?Tyr?Cys?Asn?Ser?Tyr?Val?Gly?Thr?Cys?Ala?Tyr?Met?Ser
195 200 205
Pro?Glu?Arg?Phe?Asp?Ser?Ala?Ala?Gly?Glu?Asn?Ser?Asp?Val?Tyr?Ala
210 215 220
Gly?Asp?Ile?Trp?Ser?Phe?Gly?Val?Met?Ile?Leu?Glu?Leu?Phe?Val?Gly
225 230 235 240
His?Phe?Pro?Leu?Leu?Pro?Gln?Gly?Gln?Arg?Pro?Asp?Trp?Ala?Thr?Leu
245 250 255
Met?Cys?Val?Val?Cys?Phe?Gly?Glu?Pro?Pro?Arg?Ala?Pro?Glu?Gly?Cys
260 265 270
Ser?Asp?Glu?Phe?Arg?Ser?Phe?Val?Asp?Cys?Cys?Leu?Arg?Lys?Glu?Ser
275 280 285
Ser?Glu?Arg?Trp?Thr?Ala?Ser?Gln?Leu?Leu?Gly?His?Pro?Phe?Leu?Arg
290 295 300
Glu?Ser?Leu
305
Claims (10)
1. Arabidopis thaliana plant type controlling gene BUD1 encoded protein matter, it has the aminoacid sequence shown in the Seq ID No.2.
2. coding claim 1 described proteinic gene.
3. gene according to claim 2, it has the nucleotide sequence shown in the Seq ID No.1.
4. plasmid that contains claim 2 or 3 described genes.
5. carrier that is used for expression of plants that contains the described plasmid of claim 4.
6. host cell that contains the described carrier of claim 5.
7. one kind contains the described host cell of claim 6, and this cell is intestinal bacteria.
8. one kind contains the described host cell of claim 6, and this cell is an Agrobacterium.
9. one kind contains the described host cell of claim 6, and this cell is a vegetable cell.
10. the method for the plant type that cultivates plants comprises with the described expression vector transformed plant cells of claim 5; With the plant transformed cell culture is become plant.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02144344 CN1216908C (en) | 2002-10-10 | 2002-10-10 | Arabidopsis thaliana BVD1 gene and its use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02144344 CN1216908C (en) | 2002-10-10 | 2002-10-10 | Arabidopsis thaliana BVD1 gene and its use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1488644A CN1488644A (en) | 2004-04-14 |
| CN1216908C true CN1216908C (en) | 2005-08-31 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN 02144344 Expired - Fee Related CN1216908C (en) | 2002-10-10 | 2002-10-10 | Arabidopsis thaliana BVD1 gene and its use |
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| Country | Link |
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| CN105713076B (en) * | 2014-12-01 | 2019-07-09 | 中国农业大学 | One kind CAD1 albumen relevant to plant tip advantage and its relevant biological material and application |
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