CN110408618A - Caragana intermedia inducible promoter CiNAC071 and its application - Google Patents

Abstract

The present invention relates to biological gene engineering fields, in particular to inducible promoter CiNAC071 and its application, CiNAC071 promoter is induced by ABA and GA, inhibit promoter activity under ABA processing, promotes promoter activity under GA processing, can be used for the building of transgene expression vector.The research of gene function is more advantageous to using inducible promoter.

Description

Caragana intermedia inducible promoter CiNAC071 and its application

Technical field

The present invention relates to a kind of inducible promoter CiNAC071 and its applications, start in particular to CiNAC071 The activity inducement of son promoter under ABA processing and under GA processing.CiNAC071 promoter is induced by ABA and GA, can be used for turning The building of expression vector.The research of gene function is more advantageous to using inducible promoter.

Background technique

Plant growth and development is the process that different genes are orderly expressed and acted synergistically on time, space.Gene is whole Switch, expression pattern and gene expression abundance in a growth course etc. are by accuracy controlling.The expression regulation of plant gene can Be divided into transcription before, transcription, transcription after, translation, translation after etc. 5 different levels, wherein the regulation on transcriptional level is most main It will be with the most key link.It is mainly by regulatory transcription regulatory factor and promoter that gene regulates and controls at transcriptional level In conjunction with whether, strong and weak variation etc. realizes.General genetically modified plants are obtained in such a way that T-DNA is inserted into Plant Genome , if transgene is closely related with plant growth and development, growth of transgenic plants will be caused to change, with As for being difficult to obtain transgenic homozygous body seed, phenotypic analysis can not be carried out.It is therefore desirable to which inducible promoter, is turning base Because in the screening of plant, which is not expressed, when studying gene function, which is induced by inducer.Meanwhile The case where expression quantity is too low in transgenic plants there are foreign gene, is unfavorable for gene functional research, is started using induction type Son is more advantageous to the research of gene function.Therefore, inducible promoter is significant in genetic engineering.

NAC transcription factor is that plant is distinctive, one of maximum transcription factor family.NAC transcription factor family is by initial ATAF (the Arabidopsis of the NAM (No apical meristem), arabidopsis that are found in morning glory Transcription activation factor) and CUC (cup-shaped cotyledon) composition.The N of the family protein End has highly conserved structural domain, is made of 150 or so amino acid sequences, as DNA binding structural domain；C-terminal phase To can be changed, as transcription regulating region, it can activate or inhibit to transcribe；Part NAC transcription factor contains transmembrane domain.NAC The DNA binding structural domain of transcription factor is made of five subdomains of A-E, and wherein substructure A may participate in the formation of dimer, The positively charged combination for participating in DNA of subdomain C and D, and subdomain B and D determine the diversity of NAC transcription factor.Turn in NAC Record the factor upstream promoter sequence, containing there are many regulation plant metabolism, growth and development and stress response regulation cis element, But its Regulation Mechanism is not also completely clear.In recent years, depositing for NAC transcription factor family gene is found in many species , such as have 117 NAC genes in arabidopsis (Arabidopsis thaliana), have 151 in rice (Oryza sativa) A NAC gene has 152 NAC genes in soybean (Glycine max), has 97 in clover (Medicago truncatula) NAC gene.

The research of arabidopsis NAC transcription factor family gene is more detailed, according to the structure and function of known NAC gene to portion Gene is divided to rename, with NST (NAC secondary wall thickening promoting factor) and SND The NAC transcription factor of (secondary wall-associated NAC domain protein) name participates in time of arabidopsis Grade wall growth, such as AT3G61910, AT2G46770, AT1G28740, AT4G28500；With NTL (NAC with Transmembrane motif 1-like) and NTM (NAC with transmembrane motif) name NAC transcription because Son have transmembrane domain motif, as AT1G01010, AT2G27300, AT3G10500, AT3G49530, AT4G01550, AT4G35580 and AT4G01540 etc.；It is base with the NAC transcription factor that SGS (suppressor of gene silence) is named Because of the aporepressor of silencing, such as AT3G10490 and AT3G10490；With NARS (NAC-regulated seed Morphology the NAC transcription factor) named is related to the seed morphology of arabidopsis, such as AT3G15510；With CBNAC The NAC transcription factor AT4G35580 of (calmodulin-binding NAC protein) name encodes calmodulin combination NAC Albumen has the function of calmodulin Transcription inhibition；With VND (vascular-related NAC domain) and VNI (VND-interacting) name NAC transcription factor it is related with the vasculation of arabidopsis, as AT5G66300, AT5G62380, AT4G36160, AT2G18060, AT1G71930, AT1G12260 and AT5G13180 etc.；With JUB1 (JUNGBRUNNEN1) the arabidopsis gene AT2G43000 named delays senescence after being overexpressed, and enhances to abiotic stress Tolerance.

NAC transcription factor participate in plant growth and development each stage, including seed sproutings, root growth, secondary wall formation, Plant hormone signal access, leaf senile etc..Existing research shows that the salt signal that NTL8 regulation GA is mediated responds approach, adjust Seed is saved to sprout；Arabidopsis NAC1 is overexpressed plant, and compared to check plant blade, bigger, lateral root is more and fresh weight increases；Arabidopsis The formation of secondary wall in middle SND1 and NST1 transcriptional activation fibrocyte；Under Abiotic stress conditions, arabidopsis RD26 The transcription activating protein of (dehydration-induced NAC protein 26) as ABA induced gene；It is overexpressed intermediate Caragana CiNAC1 and CiNAC4 promote the elongation of arabidopsis main root and the increase of lateral root number；Caragana intermedia CiNAC1 gene Also promote the aging of transgenic arabidopsis blade；Mahmood etc. studies have shown that ANAC032 promote arabidopsis aging while, In Under the stress conditions such as sucrose, salt, oxidation, inhibit the synthesis of Arabidopsis anthocyanin；Arabidopsis ANAC046 can promote arabidopsis leaf green Element degradation and leaf senile.

In addition to this, NAC transcription factor also assist in plant to the abiotic stress such as arid, salt, hot and cold, mechanical damage and The drought-resistant and saline-alkaline tolerance of NAC1 enhancing rice is overexpressed in the biotics response such as disease, insect pest, in rice；Mistake in rice Expression OsNAC6 can not only enhance anti-dehydrating and salt stress tolerance, and enhance disease resistance；Caragana intermedia CiNAC3 and CiNAC4 improves transgenic arabidopsis to the tolerance of salt stress；Arabidopsis ATAF1 lowers drought stress response base The expression of cause；Banana MusaSNAC1 can remove the H of guard cell₂O₂, cause stomata to be closed, enhance the drought resistance of banana；Cross table Up to crinosity tomato ShNAC1 enhancing plant to cold, arid, salt sensibility, meanwhile, also promote dark and Salt treatment blade to decline Always.All these are research shows that NAC transcription factor has important work in the fermentation such as plant growth and development and stress-tolerance regulation With.

The expression of eukaryotic gene is adjusted by the promoter for being located at transcriptional start point upstream, it is the knot of RNA polymerase II Coincidence point.The beginning and accuracy of promoter regulation genetic transcription are the centers of transcriptional control.A variety of promoter sequences are compared Analysis, it is found that their basic structural feature is, at sequence -20bp to -30bp, there is TATA sequence, in -70bp to -78bp , there is the area CAAT at place.Meanwhile some promoters also contain the special cis-acting elements different from other promoters.Promoter master It is divided into: composing type, tissue specificity and inducible promoter.Constitutive promoter is also known as non-specific promoter, in plant Growing in each stage different tissues position has expression, does not have time and Space-time speciality, such as cauliflower mosaic virus (CaMV) 35S promoter；Tissue-specific promoter's energy controlling gene is in the specific histoorgan expression of plant, such as soybean SbPRPI gene promoter is mainly that one kind is activated by signal stimulus in root development system expression inducible promoter, thus A kind of promoter for driving downstream gene expression can be divided into the starting of abiotic stress induction type according to the difference of inducible factor Son, biotic inducible promoter and hormone inducible promoter.The promoter element for analyzing gene is conducive to understand gene Expression pattern.

Promoter can classify according to the difference of function are as follows: constitutive promoter, tissue-specific promoter and induction type open Mover.Inducible promoter, the "on" and "off" of inducer gene expression capable of fast starting.It could start in the presence of inducer Gene expression, after removing inducer, gene expression is closed quickly, thus artificially can accurately and fast control gene Expression.

Caragana intermedia (Caragana inermedia) belongs to the perennial shrub of pulse family Caragana.Root system is more developed, Nearly 5 meters of subterraneous root, stem uprightly or obliquely, 1-2 meters of plant height；Pinnate compound leaf, leaflet 12-16 piece, obovate or ellipse are more There is coat；Caragana intermedia starts to grow mid-April, and the florescence is mid-May, and corolla butterfly, yellow, fruiting period is June, seed For kidney shape, there are two kinds of colors of pale green brown and yellowish-brown.It is extensive in Area distributions such as China Inner Mongol, Shanxi, Ningxia and Shaanxi, It is the excellent plant that arid, Desert Area check winds and fix drifting sand and conserve water and soil with drought-enduring, high temperature resistant, saline-alkali tolerant and the characteristics such as cold-resistant Object；In addition, Caragana intermedia is also used as feed, fuel, papermaking, fertilizer and plate raw material, while also there is medical value. The present invention clones from Caragana intermedia and obtains inducible promoter CiNAC071, inhibits promoter activity, In under ABA processing GA processing is lower to promote promoter activity.CiNAC071 promoter is induced by ABA and GA, can be used for the structure of transgene expression vector It builds.The research of gene function is more advantageous to using inducible promoter.

Summary of the invention

The purpose of the present invention is to provide a kind of Caragana intermedia inducible promoter CiNAC071 and its applications, that is, provide It is a kind of it is new can in plant tissue inducible promoter CiNAC071.

The purpose of the present invention is what is be achieved through the following technical solutions:

In a first aspect, the present invention provides a kind of nucleotide CiNAC071, sequence is as shown in SEQ ID NO.1；The sequence can It, can also be by PCR and digestion method from the genome of Caragana intermedia to be synthesized by artificial synthesized method It is cloned.

Second aspect, the present invention provide one group of primer pair for being used to expand the nucleotide CiNAC071, the primer Sequence is as shown in SEQ ID NO.2, SEQ ID NO.3.

The third aspect, the present invention provide a kind of recombinant vector, include the nucleotide CiNAC071, such as the company in embodiment Connect the pCAMBIA1305.2 carrier etc. of nucleotide CiNAC071.

Fourth aspect, the present invention provide a kind of transformant, include the nucleotide CiNAC071, what the transformant used Host is Agrobacterium.

5th aspect, the present invention provide a kind of purposes of the nucleotide CiNAC071 as promoter, and CiNAC071 is opened Mover is induced by ABA and GA.

Preferably, the purposes is specially nucleotide CiNAC071 as target gene in inducible promoter driving plant The application of expression inhibits downstream gene activity under ABA processing, promotes downstream gene activity under GA processing；The plant packet Include Caragana intermedia, arabidopsis.

Preferably, the nucleotide CiNAC071 is specific as the application of promoter destination gene expression in driving plant To impregnate transfection plant tissue with the recombinant vector, or with transformant direct transfection plant tissue.

Preferably, the purposes is specially application of the nucleotide CiNAC071 as promoter in plant gene function.

6th aspect, the present invention provide the preparation method of nucleotide CiNAC071 a kind of, including artificial synthesized method or Biological cloning method；

Preferably, the biological cloning method specifically:

Step 1 extracts Caragana intermedia DNA；

Step 2, using the DNA as template, successively expanded with the Outside primer, inner primer；

The amplified production is carried out agarose gel electrophoresis detection to get nucleotide CiNAC071 by step 3.

CiNAC071 promoter of the invention has the property that A) in the expression feelings of Caragana intermedia and arabidopsis tissue Condition is very clear；B) inhibit promoter activity under ABA processing, promote promoter activity under GA processing.

The present invention uses molecular cloning method for the first time, and separation obtains new CiNAC071 from Caragana intermedia genome Promoter, and it is confirmed by transgenic experiments as inducible promoter, CiNAC071 promoter is induced by ABA and GA, The building that can be used for transgene expression vector, the research of gene function is more advantageous to using inducible promoter, to reach The object of the invention.

Detailed description of the invention

Fig. 1: CiNAC071 promoter clone and expression vector digestion verification: figure A: chromosome walking method expands for the first time CiNAC071 promoter schemes B: second of chromosome walking method and expands CiNAC071 gene promoter, and swimming lane 1,2,3 respectively indicates First round PCR, the second wheel PCR, third round PCR；Scheme C:CiNAC071 gene promoter clone, 1,2 respectively indicate identical PCR Product；Figure D: digestion verification Pro_CiNAC071: GUS, swimming lane D: plasmid control；000 bp DNA marker of M1:DL5；M:DL2 000bp DNA marker。

The analysis of Fig. 2: CiNAC071 promoter cis element.

Fig. 3: Pro_CiNAC071: the histochemical stain of GUS transgenic line and wild-type plant: to Pro_CiNAC071:GUS Transgenic line and each histochemical stain of wild-type plant different growing stage, a-h are the histochemical stain of seedling period, i- R is plant growth 40 day maturity period each histochemical stain.

Fig. 4: MeJA, ABA and GA handles lower Pro_CiNAC071: the histochemical stain analysis of GUS genetically modified plants.

Specific embodiment

The present invention is further described in detail With reference to embodiment, and the embodiment provided is only for explaining The bright present invention, the range being not intended to be limiting of the invention.Experimental method in following embodiments is unless otherwise specified Conventional method.The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.

Embodiment 1.CiNAC071 promoter clone

The promoter for obtaining CiNAC071 gene is cloned using Genome Walking kit.According to gene ORF sequence, Design obtains first round promoter and clones special primer SP1, SP2 and SP3, referring to table 1.The primer and four kinds in kit it is general Primer carries out hot asymmetric PCR reaction using Caragana intermedia gDNA as template, obtains gene flank sequence by nest-type PRC three times Column.Clear and special amplified band, connects pEASY-T1Simple cloning vector after recycling third time PCR amplification, recombinates matter It is sequenced after grain conversion to Escherichia coli.Analyze sequencing result can the splicing of isogenic open reading frame sequence it is errorless, then it is assumed that The sequence is the promoter of gene.If the first round expands the sequence for not obtaining ideal length, the second wheel amplification can be carried out.

The primer of second wheel promoter amplification, is designed on the basis of the first round obtaining sequence.It is still three special in the same direction Different primer 2 nd-SP1,2nd-SP2 and 2nd-SP3 obtain second of promoter clone's using Caragana intermedia gDNA as template Purpose band.After sequencing analysis, the sequence is errorless with first round amplification acquisition sequence assembly, then the sequence assembly expanded twice Getting up is the promoter sequence of the gene.

Meanwhile the primer p071-sense and p071-anti of amplification promoter overall length are designed with splicing result, with intermediate brocade Chicken gDNA is template, expands promoter sequence.

PCR program

After PCR amplification, target fragment is recycled using TIANGEN Ago-Gel DNA QIAquick Gel Extraction Kit.

1 list of primers of table

Embodiment 2, Caragana intermedia CiNAC071 promoter connect pCAMBIA1305.2 expression vector

The restriction enzyme site Hind III and Nco I for including according to special primer, by empty carrier pCAMBIA1305.2-GUS digestion After processing, it is connect with target fragment by In-fusion mode.Linked system is as follows:

Each substance is centrifuged after mixing, 37 DEG C of water-baths 15min, 50 DEG C of water-bath 15min.It converts to E. coli competent DH5 α is verified using double digestion and bacterium colony PCR.Pro_CiNAC071: after GUS recombinant plasmid expression vector constructs successfully, convert large intestine Bacillus is extracted recombinant plasmid, is verified using double digestion and bacterium colony PCR.Using the small extraction reagent kit of plasmid by recombinant plasmid from It is extracted in Escherichia coli, electrotransformation to Agrobacterium tumefaciems GV3101.Use dip method transfected wild-type arabidopsis, hygromycin selection It obtains and is overexpressed plant.

Embodiment 3, Caragana intermedia CiNAC071 promoter clone and promoter drive gus reporter gene expression vector structure It builds

CiNAC071 gene ORF full length sequence is obtained from Caragana intermedia transcript profile database, in known array, if Three reverse primers 071-SP1,071-SP2 and 071-SP3 are counted, it is anti-by three-wheel PCR using Caragana intermedia gDNA as template It answers, amplification obtains the promoter (Figure 1A) of CiNAC071 gene, recycles two distinguished sequences of third round PCR product.Through being sequenced Analysis, length are in 1000bp between 1500bp, and the brighter sequence of electrophoretic band is the promoter of CiNAC071 gene, should Sequence length is 1246bp, which is used to analyze not enough whole cis elements of CiNAC071 gene promoter, because This, continues to do second of promoter amplification.

In the sequence basis generated after first time PCR sequence is with the connection of CiNAC071 gene ORF full length sequence, design three Reverse primer 071-2nd-SP1,071-2nd-SP2 and 071-2nd-SP3.Again using Caragana intermedia gDNA as template, warp Three-wheel PCR reaction is crossed, second of product (Figure 1B) expanded of CiNAC071 gene promoter is obtained.It this time expands, equally has Two specific bands, are analyzed through sequencing result, length be located at 750bp to the sequence between 1000bp, more bright can be with first Secondary extension increasing sequence splicing, thus it is speculated that it should be the partial sequence of CiNAC071 gene promoter.

Promoter twice is expanded to the sequence assembly obtained, constitutes the promoter sequence of CiNAC071 gene, altogether 2107bp.According to CiNAC071 gene promoter sequence, clone-specific primers p071-sense and p071-anti are designed.With Caragana intermedia gDNA is template, and under the catalysis of PrimeSTAR enzyme, clone obtains CiNAC071 gene promoter (Fig. 1 C). By pCAMBIA1305.2 carrier using Hind III and I restriction endonuclease of Nco linearisation after, under In-fusion system with PCR product Connection building Pro_CiNAC071: GUS expression vector.Hind III and I double digestion qualification figure of Nco generate purpose after digestion referring to Fig. 1 D Genetic fragment and linearized vector, show Pro_CiNAC071: the success of GUS expression vector establishment.

Gene promoter sequence predicts various response elements included in the promoter through PlantCARE on-line analysis, Thus infer regulating and controlling effect of the gene in each signal path.Analyze the element of taking advantage of a situation of the sequence promoter: eucaryote is opened The common transcriptional elements CAAT-box and TATA-box of mover；Plant light response element ATC-motif, ACE, ATCT-motif, G- Box etc.；Separate living tissue expresses CAT-box；Plant hormone methyl jasmonate response element CGTCA-motif, gibberellin response member Part P-box, TATC-box, abscisic acid response element ABRE；Stress response element TC-rich repeats, low temperature stress response Element LTR；Zeins metabolic regulation element O2-site etc..Speculate that CiNAC071 gene may participate in from these elements The biochemical reactions such as hormone regulating and controlling and stress response.

Embodiment 4, Pro_CiNAC071: GUS Transgenic plant tissue chemical staining

Successful Pro will be constructed using flower-dipping method_CiNAC071: GUS plasmid transfection arabidopsis wild-type plant, hygromycin selection Obtain transgene positive plant.It is control, histochemical stain of the observation T2 for plant different growing stage with wild-type plant Situation.Select four time Vertex Colorings, be for the first time seed sprout 48h, dyeing be concentrated mainly on cotyledon and it is lower with axis (Fig. 3 a and e)；It is that seed sprouts 5d for the second time, dyeing is concentrated mainly on cotyledon, hypocotyl, at petiole (Fig. 3 b and f)；Third time is seed 10d is sprouted, dyeing, which is concentrated mainly at cotyledon vein, newborn true leaf blade, hypocotyl and small part root, (Fig. 3 c, Fig. 3 d, to be schemed 3g and Fig. 3 h).4th time is plant growth 40d, main to the lotus throne leaf of plant, flower, stem and mature fruit pod staining analysis, dyeing Vein, calyx, column cap, fruit pod top end are concentrated on, and stem absolutely not dyes.Wild-type plant is in growth different phase and not With tissue without dyeing (Fig. 3 i-r).Studies have shown that the gus reporter gene of CiNAC071 promoter driving is mainly in leaves of plants The expression of the histoorgans such as piece, hypocotyl and calyx.

Embodiment 5, Pro_CiNAC071: GUS genetically modified plants HORMONE TREATMENT histochemical stain

There are 5 ABA response elements, 4 MeJA responses in CiNAC071 promoter in the higher cis element of the frequency of occurrences Element and 2 GA response elements.In order to analyze response of these elements to plant hormone, above-mentioned HORMONE TREATMENT is used Pro_CiNAC071: after GUS genetically modified plants, do histochemical stain analysis.As shown in figure 4, Pro_CiNAC071: GUS genetically modified plants With more no significant change before handling after MeJA is handled.But after ABA is handled, staining signals increase with ABA concentration and are subtracted It is weak, but enhance with the increase of GA concentration, shows that CiNAC071 promoter is able to respond ABA and GA, be inducible promoter.

SEQUENCE LISTING

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<120>Caragana intermedia inducible promoter CiNAC071 and its application

<130>Caragana intermedia inducible promoter CiNAC071 and its application

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<170> PatentIn version 3.3

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<213> Caragana inermedia

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tgtgaggtga cttgctgctg tgctgcagcc tcatacagag cagcaggctc taatttagct 120

agtactacct atatgttttt agaatatttt attccaaata aaagatgagc atatctattc 180

cttgaagcat gaatcacata gggggtaacg tgtctgactt gccaaaaaaa gcaaatgcgt 240

tgcatctcag aaaagatatt tcaagagaaa tttatttgtt cagtcataaa aaaaaaagag 300

agatttattt gtctttttca attactaata ttaaatagac aattatttat taaaaaaagt 360

attagtacgt ttatatataa aatccaacaa aaaaaaaatg ttaaatgaat acgtgtgcat 420

ctcacactat ctcttgcctc ggctgcttca caccttcacc attctcgatc taacgcagac 480

catcttctct gaagggccta tttgcatttt ttttcatctt cctccatcgc cgaccacttc 540

ttttggccga agttgctgtc aaccgtttcc cttcatcttc ccttcctcct ttctctgttt 600

ttttttctca tctttaccaa ctatggttga tgcttctgcc cctgcaactt gatcagcaac 660

gtgcggctcc ccctctgtac agcactccgg cgccgccatt tgcttcacct ctggtggcat 720

tgagcttacc cgggcaaatc cgtccgatcg gaaccgtcac ccgaccaaac cgtcaccggc 780

caaaccgtgc aagaaaacgg acggaattgg gccacatatt acagcccacg gtttgggccg 840

gattagagtt ttttggcccg aaaccgatcc ggcccgaacc atgtccagcc ctactcatca 900

tcatcatcat tcatgatgac gctttgataa ttgaaaggca atttaacgaa aggtcatact 960

aacaagtgct ttaaagacat tgtttaagga ttctaaataa ggtaaatact catttaaaaa 1020

ccatgtttat ctctttccaa cgtattaaat acacaacttc tcataaaaac ttactttttt 1080

ggacttctta aacaatactc ttagggcact cattagtatt ttccttaatg aaattagagg 1140

catataatcc tctgcttttg ttagtgattg aaggcaagct aatgttgacg tccattcttt 1200

aactgagatg caaaggaaga agctaatgtt aattaagaac ccaaaacgca tgaaataaat 1260

aagccataaa ataacggtcg caaattattg gaattggcgg gaagtaaact aaactttgta 1320

ggtgtgagct ggttggagtg gtgggtgggc cattttgctt tggaattttt ggtgcactcg 1380

tgacaagtaa tctcaaactt tcggttcagt gtgctcaatt ttggactgtt ggttcccttc 1440

cccaaatgct ataaatacca caccatcact gacctatgtt tttactctat agtctcttgt 1500

cttcatcatc tcaaattaaa gcactcacag ttccatcacc atcacactca cttctgaact 1560

tgtttattct aatttctggc ctttgattag ctgattagct ccctcctttt ggaatcttgt 1620

gataaaattt ctgatttgtt atttttgttt tgtatatttg ggggtcaaaa gttggtaatt 1680

taataattgg taacttctga gtattactaa gttaatatca attctttttg ttccagcttt 1740

tcagggactt ttgaatatta aggggaggtg ggttggggtg ggatagctgc cataagacac 1800

atctaattac ttcaagacga tgatccttgc attcaatgga tgcactttgg ggaagtgcca 1860

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Claims (9)

1. a kind of nucleotide CINAC071, which is characterized in that its sequence is as shown in SEQ ID NO.1.

2. one group for expanding the primer pair of nucleotide CINAC071 according to claim 1, which is characterized in that the primer Pair sequence as shown in SEQ ID NO.2, SEQ ID NO.3.

3. a kind of recombinant vector, which is characterized in that include nucleotide CINAC071 according to claim 1.

4. a kind of transformant, which is characterized in that include nucleotide CINAC071 according to claim 1.

5. a kind of purposes of nucleotide CINAC071 according to claim 1 as promoter.

6. a kind of preparation method of nucleotide CINAC071 according to claim 1, which is characterized in that including artificial synthesized Method or biological cloning method.

7. the preparation method of nucleotide CINAC071 according to claim 6, which is characterized in that the biological cloning method tool Body are as follows:

Step 1 extracts Caragana intermedia DNA；

Step 2, using the DNA as template, successively expanded with the Outside primer, inner primer；

The amplified production is carried out agarose gel electrophoresis detection to get nucleotide CiNAC071 by step 3.

8. a kind of application of nucleotide CINAC071 according to claim 1 in plant gene function research.

9. the application in plant gene function research according to claim 8, the promoter are induced by ABA and GA, thus Control the expression of downstream gene.