CN109762815A - One kind specifically expressed promoter PCHF17 and its application in Rice Anther and pollen - Google Patents

Abstract

The invention discloses one kind specifically expressed promoter PCHF17 and its applications in Rice Anther and pollen, are related to genetic engineering and molecular biology field.Promoter PCHF17 of the present invention has nucleotide sequence shown in SEQ ID NO.1, the present invention further provides the expression vector containing promoter PCHF17, expression casette, engineering bacteria or cell line.The present invention also provides the primer pairs of amplification anther and pollen specific promoter.Anther and pollen-specific promoter of the invention is rice endogenous gene, it is highly beneficial to paddy gene engineering, foreign gene can be driven accurate in anther and specific expressed and expression in pollen, provide driving foreign gene new method specific expressed in Rice Anther and pollen.

Description

One kind specifically expressed promoter PCHF17 and its application in Rice Anther and pollen

Technical field

The present invention relates to genetic engineerings and molecular biology field, specifically, being related to Rice Anther and pollen specific Promoter PCHF17 and its application.

Background technique

Transcriptional control is one of principal mode of control of plant gene expression, by cis-acting elements and trans-acting factor Coordinate to complete.Promoter is one of most important cis element in plant gene transcription regulation, is normally at gene 5 ' and holds upstream, It is identification and the binding site of RNA polymerase and some trans-acting factors.Promoter mainly includes core promoter Qu Yuzhuan Record two functional areas of control region.Core promoter area is the most short promoter fragment of starting transcription, and generally 40nt is one section of quilt DNA sequence dna RNA polymerase family I, II and III identification and combined.This region includes some important function element, Neng Gouzhun True positioning transcriptional start point and direction are the bases of gene expression regulation.Transcription regulating region is located at the upstream of core promoter (or downstream), can be combined with special transcription factor to the space-time of transcription and it is strong and weak play regulating and controlling effect, such as enhancer and Silencer etc..The expression pattern of further investigation promoter not only improves the expression and regulation mechanism and biological function for understanding gene, Facilitate the expression of control foreign gene again.

Promoter can be divided into constitutive promoter, inducible promoter and space-time specific promoter etc. by its expression way Three classes.Constitutive promoter promotor gene can transcribe in all or most of tissues, and gene expression is made to have space-time duration With expression quantity constancy.The 35S promoter of tobacco mosaic virus (TMV), the Actin promoter of rice and the Ubiquitin of corn are opened Mover belongs to constitutive promoter.Constitutive promoter is widely used in the genetic engineering research of plant, is used for target base Cause, such as pest-resistant and anti-herbicide gene overexpression.Inducible promoter can open under the stimulation of certain physically or chemically signals Move or greatly improve gene expression.They have the sequential structure of enhancer, silencer or similar functions, and have apparent single-minded Property.Inducible promoter can be divided into photoinduction promoter, heat shock promoter, low temperature induction according to the difference of inducement signal and open Mover, drought-induced promoter, wound-induced promoter, hormone induction promoter etc..Space-time specific promoter is only specific Growth phase or position in promotor gene expression.Tissue-specific promoter is one kind of space-time specific promoter, is only existed Start expression in specific cells, tissue or organ.It is controlled in the genetic transformation of plant with the promoter of organizing specific expression The expression of target gene can more effectively avoid such as can be reduced composing type table using the potential negative interaction of constitutive promoter bring Up to increased metabolic burden, reduction Safety of GM Food risk and to the adverse effect with environment, and reuse identical The gene silencing etc. that promoter causes.The type of current developed rice tissue specific promoter is varied, root, stem, Leaf, seed and fruit etc. have almost had been found that organized specific expressed promoter in various tissues.

Male sterility is the basis of heterosis utilization, is the core technology of hybrid rice industry, Study On Rice fertility tune The mechanism of control has great theoretical and practical significance.Anther is that rice generates mature male gametophyte (i.e. pollen or microspore) Organ.In the early stage of anther development, there are a kind of primary sporogenous cell, which passes through a series of divisions and differentiation, produces Raw tapetum and pollen mother cell.Tapetum provides nutrition, including various enzymes and non-enzymatic albuminoid for the development of male gametophyte Matter, and participate in the synthesis of exposore.Pollen mother cell then carries out meiosis, generates tetrad.It is unicellular in tetrad Effect through tapetum secretion callosity enzyme separates Haploid production microsporinite.Monoploid microsporinite is further developed, to evening The big vacuole in center is formed when the phase.Central big vacuole squeezes nucleus to cell edges (referred to as mid-late uninucleate stage), forms cell Polarity.This cell polarity promotes microsporinite to carry out primary unequal mitosis, generates a big vegetative cell and one A small reproduction cell.Small reproduction cell is completely contained in big vegetative cell, and cell is special existing in formation cell As.Reproduction cell then carries out second of mitosis, generates two spermatids, forms mature pollen grain.Therefore, screening and Determine that Rice Anther and pollen-specific promoter by for the genetic engineering of rice, are especially prevented in sterility changing and transgenosis drift Control etc. provides new selection.

Summary of the invention

The object of the present invention is to provide a kind of plant anthers and pollen specific promoter and its application.

A kind of plant anther provided by the invention and pollen specific promoter PCHF17, to include

1) nucleotide sequence shown in SEQ ID No:1,

Or 2) one or more nucleotide are substituted, lack or added in the nucleotide sequence shown in SEQ ID No:1 And the nucleotide sequence as derived from 1) with same anther and pollen specific startup function；

Or 3) the sequence complementary with nucleotide sequence shown in SEQ ID No:1.

Wherein, the nucleotide sequence as derived from 1) described in 2), the nucleotides sequence described in 1) is classified as to be had compared with 1) 70% or more identity, 80% or more identity, 85% or more identity, 90% or more identity, 95% or more identity, 98% or more identity or 99% or more identity, and the nucleotides sequence with same anther and pollen specific promoter function Column.

Those skilled in the art can easily identify and utilize and plant anther and pollen specific according to identical purpose Therefore the DNA molecular of promoter PCHF17 nucleotide sequence complementation has promoter activity and under strict conditions can be with this hair Bright promoter sequence or the DNA sequence dna of its segment hybridization are included in the invention.Wherein, the nucleotide sequence is complementary, refers to It can hybridize under strict conditions with PCHF17.

It is more than to hybridize with other sequences (as at least that stringent condition, which refers to that probe will be hybridized to detectable degree with its target sequence, 2 times of backgrounds) condition.Stringent condition has sequence dependent, and different because of the difference of the other conditions of experiment.Pass through control The stringency of hybridization and/or wash conditions, can identify the target sequence complementary with probe 100% (same to source detection).Selectively, Adjustable stringent condition is to allow some sequence mismatch, so that detecting the similitude (heterologous detection) of lower degree.In general, Probe length is no more than 1000 nucleotide, is preferably shorter than 500 nucleotide.

Typically, stringent condition is that salinity is lower than about 1.5M Na ion at pH value 7.0-8.3, typically about 0.01-1.0M Na ion concentration (or other salts), temperature are right at least about 30 DEG C of short probe (such as 10-50 nucleotide) At least about 60 DEG C of long probe (such as more than 50 nucleotide).It also can get stringent item by adding destabilizing agent such as formamide Part.Low stringency condition, it may for example comprise molten in 30-35% formamide, 1M NaCl, the buffering of l%SDS (dodecyl sodium sulfate) 37 DEG C of hybridization in liquid, 1 × the 50-55 DEG C of washing into 2 × SSC (20 × SSC=3.0M NaCl/0.3M trisodium citrate).In Spend stringent condition, it may for example comprise 40-45% formamide, 1.0M NaCl, l%SDS buffer solution in 37 DEG C hybridization, 0.5 × 55-60 DEG C of the washing into 1 × SSC.High stringency, it may for example comprise in 50% formamide, 1M NaCl, l%SDS Buffer solution in 37 DEG C hybridization, in 0.1 × SSC 60-65 DEG C washing.Optionally, washing buffer can be containing about The SDS of 0.1%-1%.Hybridization time is generally less than about 24 hours, generally about 4-12 hours.

Especially typically the function of post-hybridization washing, key factor are the ionic strength and temperature of final washing solution. For DNA-DNA hybrid, Tm can be from the equation of Meinkoth and Wahl (Anal Biochem, 1984,138:267-284) Estimation: Tm=81.5 DEG C+16.6 (logM)+0.41 (%GC) -0.61 (%form) -500/L；Wherein M is univalent cation Molar concentration, %GC are the percentage of guanylic acid and cytidylic acid in DNA, and %form is that formamide is hybridizing Percentage in solution, L are length of the hybrid in base-pair.Tm is that 50% complementary target sequence hybridizes with complete pairing probe Temperature (under defined ionic strength and pH).Every 1% mispairing needs Tm to reduce about l DEG C；Therefore, Tm hybridizes and/or washes The condition of washing can be conditioned to hybridize with the sequence of required identity.For example, if the sequence sought has >=90% identity, Tm can reduce by 10 DEG C.Generally, the stringent condition of selection is less than about 5 DEG C of thermal melting point (Tm) of particular sequence, and It is complementary under defined ionic strength and pH.But high stringency can be using lower than thermal melting point (Tm) 1,2,3 Or 4 DEG C of hybridization and/or washing；Moderate stringency can be using miscellaneous lower than 6,7,8,9 or 10 DEG C of thermal melting point (Tm) It hands over and/or washs；Low stringency conditions can apply hybridization of 11,12,13,14,15 or 20 DEG C lower than thermal melting point (Tm) And/or washing.Those of ordinary skill in the art can understand that hybridization and/or the condition of washing solution become with the variation of stringency Change, and the hybridization of this equation calculation of application and cleaning compositions and required Tm.If required extent of mismatch makes Tm lower than 45 DEG C (aqueous solution) or 32 DEG C (formamide solution), preferably increase SSC concentration is to be able to use higher temperature.The guide of nucleic acid hybridization See Tijssen (1993) biochemistry and the nucleic acid probe hybridization of Molecular Biology Lab's technology one, part i, the 2nd chapter (Elsevier, New York)；With Ausubel et al. editor the 2nd chapter (Greene of (1995) Current Protocols method Publishing and Wiley-Interscience,New York).See Sambrook et al. (1989) molecular cloning: experiment Room handbook (second edition, Cold Spring Harbor Laboratory Press, Plainview, NewYork).

The stringent condition is preferably the solution in 6 × SSC (sodium citrate), 0.5%SDS (dodecyl sodium sulfate) In, hybridize at 65 DEG C, is then respectively washed film 1 time with 2 × SSC, 0.1%SDS and 1 × SSC, 0.1%SDS.

Expression casette of the present invention offer containing plant anther of the present invention and pollen specific promoter PCHF17, Expression vector and host cell containing the expression vector.

The expression casette is to have structural gene in the downstream connection of anther and pollen specific promoter PCHF17, adjust Section gene, structural gene antisense gene, adjust gene antisense gene or can interfere with endogenous gene expression MicroRNA gene Expression cassette.

The present invention further provides the plants containing above-mentioned anther and the expression casette of pollen specific promoter PCHF17 Object.

Expression cassette the present invention provides above-mentioned Rice Anther and pollen specific promoter PCHF17 or containing it, expression Carrier or host cell are in the application for driving foreign gene specific expressed in plant anther and pollen.

Expression cassette, expression vector the present invention provides Rice Anther and pollen specific promoter PCHF17 or containing it Or application of the host cell in prepare transgenosis plant.

The genetically modified plants are foreign gene specifically expressed genetically modified plants in anther and pollen, are preferably awarded Powder/fertility enhancing/weakening genetically modified plants, more preferably male sterility genetically modified plants.

The plant includes but not limited to rice, corn, jowar, barley, oat, wheat, grain, sugarcane, soybean, Btassica Species, cotton, safflower, tobacco, clover and sunflower.

The present invention also provides the primer pair for expanding the anther and pollen specific promoter PCHF17, the primer pairs Nucleotides sequence be classified as SEQ ID NO:2-3.

The present invention provides a kind of method for separating anther and pollen specific promoter PCHF17, to use SEQ ID The nucleotide sequence of NO:2-3 primer pair PCR amplification anther and pollen specific promoter PCHF17.

The present invention also provides driving foreign genes in anther and specifically expressed method in pollen, includes the following steps:

By Rice Anther and pollen specific promoter PCHF17 of the invention and purpose exogenous gene cloning into carrier The recombinant expression carrier of the expression cassette containing PCHF17 and purpose foreign gene is obtained, and is introduced into Plant Genome, is obtained Foreign gene specifically expressed genetically modified plants in anther and pollen.

Anther provided by the invention has the advantages that with pollen-specific promoter PCHF17

1) PCHF17 is rice DNA sequence endogeneous, and Transgene-safty risk is extremely low.

2) PCHF17 can drive foreign gene anther with it is specific expressed in pollen, expression is accurate.

3) the present invention provides a kind of new methods that driving foreign gene is specific expressed in anther and pollen.

Detailed description of the invention

Fig. 1 is the recombinant expression carrier 1300gus- of anther and pollen specific promoter PCHF17 in embodiment 2 PCHF17 constructs flow chart PCHF17 map.

Fig. 2 is that 11 Rice Anthers and pollen GUS stained photographs are spent in non-transgenic.

Fig. 3 is 1300gus-PCHF17 transgenic paddy rice anther and pollen GUS stained photographs in embodiment 4.

Fig. 4 is 1300gus-PCHF17 transgenic paddy rice gynoecium stained photographs in embodiment 4.

Fig. 5 is 1300gus-PCHF17 transgenic paddy rice glume stained photographs in embodiment 4.

Fig. 6 is 1300gus-PCHF17 transgenic paddy rice root dyeing photo in embodiment 4.

Fig. 7 is 1300gus-PCHF17 transgenic paddy rice stem stained photographs in embodiment 4.

Fig. 8 is 1300gus-PCHF17 transgenic paddy rice leaf stained photographs in embodiment 4.

Specific embodiment

The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Without departing substantially from spirit of that invention In the case where essence, to modifications or substitutions made by the method for the present invention, step or condition, all belong to the scope of the present invention.

Unless otherwise specified, the conventional means that technological means used in embodiment is well known to those skilled in the art.

The acquisition of embodiment 1 Rice Anther and pollen specific promoter PCHF17

1. the extraction of oryza sativa genomic dna

Oryza sativa genomic dna is extracted using plant DNA Isolation kit (Chengdu Fu Ji Bioisystech Co., Ltd).Gene Group derives from the Fresh leaves of rice varieties OryzasativaLcv.Nipponbare.It is saved backup after the genomic DNA packing of extraction in -20 DEG C.

2.PCHF17 PCR primer design and amplification

Design of primers uses Gibson Assembly method, and amplified production is inserted into 1300GUSplus carrier (will GUSplus element be inserted into pC1300 multiple cloning sites obtain) Nco I and Hind III digestion site.Amplification PCHF17's draws Object sequence is as shown in SEQ ID NO:2 and SEQ ID NO:3.Wherein there are 15 nucleotide sequences at the end of upstream and downstream primer 5 ' and carry The corresponding link position overlapping of body, so as to Gibson Assembly connection.

PCR reaction system (100 μ L): DNA profiling: 3 μ L (50ng), KOD polymerase (be purchased from Japan mill company): 2 μ L, 10 × buffer:10 μ L, 10 μM of forward primers: 3 μ L, 10 μM of reverse primers: 3 μ L, 10 μM of dNTP:10 μ L, MgSO₄: 4 μ L, 1/ 10DMSO:20 μ L, ddH₂O:45 μ L.

PCR program: 95 DEG C of initial denaturation, 4min.94 DEG C of denaturation, 30s；50 DEG C of annealing, 30s；Extend 68 DEG C, 2min；35 Circulation.Extend 68 DEG C, 10min.

Amplified production includes the anther and pollen specific promoter PCHF17 (sequence such as SEQ ID NO:1) of 2074bp.

The recombinant expression carrier p1300gus-PCHF17 of the building of embodiment 2 promoter PCHF17

The PCR product obtained in embodiment 1 is subjected to electrophoresis, recycling 2074bp or so size in 1% Ago-Gel Band.With Nco I and Hind III double digestion carrier p1300GUSplus, linear digestion carrier is recycled.

With Lightening Cloning Kit (Jin Fusai (Beijing) Biotechnology Co., Ltd) to PCR recovery product and Linearisation p1300GUSplus empty carrier is attached, and 10 μ L systems are as follows: 2.5 μ L recovery products (50ng/ μ L), 0.5 μ L enzyme It cuts carrier (100ng/ μ L), 2.5 μ L Ligation Mix.Linker: 50 DEG C, 60min.

Take the above-mentioned 5 electroporated competent escherichia coli cell of μ L of connection product.Use primer SEQ ID NO:4 and SEQ ID NO:5 carries out bacterium colony PCR, selects positive colony sequencing verifying.Correct carrier is sequenced and is named as p1300gus-PCHF17 (map is shown in Fig. 1).P1300GUSplus carrier contains gus gene.Blue is presented in the tissue of expression gus gene after dyeing, can It is used to indicate the expressive site and intensity of promoter.

The acquisition of the rice of 3 turns of p1300gus-PCHF17 of embodiment

Take the Agrobacterium EHA105 of -70 DEG C of preservations in containing 50 μ g/mL rifampin plate streakings, 28 DEG C of cultures.Picking single bacterium It falls and is inoculated in 50mL YEB fluid nutrient medium, 28 DEG C of shaken cultivation 12-16hr of 220rpm.Take the switching of 2mL bacterium solution in 100mL In (containing antibiotic) YEB fluid nutrient medium, 28 DEG C of 220rpm shaken cultivations to OD600=0.5.It is pre-chilled 10 minutes on ice, 5000rpm is centrifuged 10min (refrigerated centrifuge is pre-chilled to 4 DEG C).Aseptic deionized water is washed 2 times (each 10mL), and 10% glycerol washes 1 It is secondary to be dissolved in 10% glycerol of 3mL.100 μ L competent cells are taken to add p1300gus-PCHF17 matter obtained in 1 μ L embodiment 1 Grain, 2.5KV are electroporated.28 DEG C of cultures, select positive colony on YEB culture plate containing kanamycin and rifampin, use P1300GUSplus vector-specific primers SEQ ID NO:4 and SEQ ID NO:5 carries out PCR verifying.

Verifying is correctly cloned, and is infected by Agrobacterium-mediated genetic transformation method and is spent 11 (Hiei Y Ohta in rice S,Komari T,Kumashiro T(1994)Efficient transformation of rice(Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T- DNA.The Plant Journal6:271-282).Through co-cultivation, the links such as screening, breaks up, takes root and obtain T₀For transgenosis children Seedling.The blade total DNA for extracting transformed plant carries out PCR positive detection, choosing using primer SEQ ID NO:6 and SEQ ID NO:7 The positive plant cultivation of the PCR that learns from else's experience verifying, self-fertility obtain T₁Generation.Take T₀Or T₁Subsequent analysis is carried out for plant.

4 transgenic paddy rice GUS staining analysis of embodiment

Prepare GUS dyeing liquor X-Gluc reaction solution (50mM phosphoric acid receives buffer, pH value 7.0, the 0.5mM potassium ferricyanide, 0.5mM potassium ferrocyanide, 0.5mg/ml X-Gluc, 20% methanol of volumn concentration, 0.1%Triton X-100), at random Choose 5 or more the transgenic positive strain obtained in embodiment 3 acquisition anther, pollen, gynoecium, glume, root, stem, leaves etc. Tissue sample is immersed in 37 DEG C of 2h in X-Gluc reaction solution or stays overnight, then sloughs tissue with 75% ethyl alcohol of volumn concentration Chloroplaset color after observe photograph.As a result as shown in Figures 2 to 4, spend 11 anther and pollen that cannot dye (figure in wild type 2), and the anther wall of transgenic paddy rice is dyed to blue (Fig. 3), has half to be dyed to blue (Fig. 3) in pollen, other gynoeciums The tissues such as (Fig. 4), glume (Fig. 5), root (Fig. 6), stem (Fig. 7), leaf (Fig. 8) cannot be colored, and show that PCHF17 promoter can Drive gus gene Rice Anther with it is specific expressed in pollen.

Although above the present invention is described in detail with a general description of the specific embodiments, On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause This, these modifications or improvements, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.

Sequence table

<110>Hainan Bo Lian paddy gene Science and Technology Ltd.

<120>a kind of specifically expressed promoter PCHF17 and its application in Rice Anther and pollen

<130> KHP181117645.4

<160> 7

<170> SIPOSequenceListing 1.0

<210> 1

<211> 2074

<212> DNA

<213>artificial sequence (Artificial Sequence)

<400> 1

cgttgtcagg cctactgggt aagtgcattt ctaggtggat tattttctca gaacttgcaa 60

ttgacatttt gttctgacct gtttctgttt gttaaacagc caaataggaa ctgaatcaga 120

gattctaact atcgatataa agcccggatg gaagaaagga accaagatta cgttcccgga 180

caaggggaat gagcagccaa accagctccc tgctgacctt gtatttgtaa tagatgaaaa 240

gccccatgat ctgtacacaa gagaaggtaa tgatctcctt gtgcaccaga agattgaatt 300

ggtggatgcg ttagcaggaa ccaccgttaa tctgaagact ctcgatggcc gggacctggt 360

gatcaagctc actgatgtgg tcacacctgg gtacgagctc gcgatcgcaa aggaagggat 420

gcctattgtc aaggagaatg ggaggagagg caatctaagg atcaagttcg acatcgtttt 480

ccccaagagg ttatcgtcag atcagcggca gaacattagg aaggttcttg gagggcaaac 540

tcagcagcag tgagcacatt ttcagaagca aatcgactgt gccatagttc ttgagataaa 600

cagactaatt cagagctctt gttcttcttc tccagttggc ttgtacacat agttattttt 660

catttggagt tgttcatctt tgatctgccc cccttcttgg aaaaatagca atgttaattg 720

atgaccacta cacgtgtaag caaaaacaca tatcattaca cttgaaatac agagtgcaat 780

gcaatgtacg tttggtgaag cccccctttt ttgtgtgaat tgctgttctt tgatttaaaa 840

caaatgaatc tattggaggt caaggcatct gttcagcatg tgaaattagc atcatcaact 900

tccttgaggg gtattgctgt agcatctaaa agaagaaaaa aatgcttctc cttggcacta 960

agacggaagt gaagggtcag ggacccaaat ccttcacata tatgtgacaa aatcacaaaa 1020

ggtagaattg tatatatgag atcttcaaag caaaaaaaca aagcttaata acatcttcag 1080

aatttcagac ccttaatgta cacaatgcat gggctgaaat gggccagaat tgcaaaatac 1140

cgcagcactt gtttgaaata taaagtgaat tgcctgtctt tcttcatcat cttaagtttt 1200

tgagttaaaa ttgttggtgc atgcaactca agagtaatat ccccacactg aattcctaag 1260

cctcaaaagg aagatttttt attttaagat aatggacatt tcattaagca aataaaagca 1320

tccggcttct gcaccaaagt gcattcaacc aagcctcaac gaatatgatg ccactaactt 1380

tatggagtac ctcctttcaa acaaaagaat ttttggagca attttgcagg aattagacta 1440

attccttcga aaatcctata aaaaattagc atccaaagga tcacctaggc cccgttcgtt 1500

tcctctgata actttggatg aagttttgga ttttcgtggc acgcttttca aactgctaaa 1560

tagtgtgttt cgtgcgaaaa ctttctctat gaaagttgtt ctaaaatatc agattaattt 1620

atttttcaag tttgtaataa ttaaaactta attaatcaca cgttattacc acctcgtttt 1680

atgtaaaaca cttaatcttc aacttcagga gattcaatca ccaccttaat gtttcacagc 1740

tactcccttc atcctggatt gctatattcc gggacagaga aaataacata tgtttttttc 1800

cctaaattat acatagttcc accatgtcat ctaaattcta aactcctttt ttttcccaga 1860

gagaaaaaaa aatctaaaat acatttcttc taccgtccat aaatagcaac ggcctcgctg 1920

catccgtggg cccacctgct ttaaccgtcg tcttctccat ttccgtttcg tggtccccgt 1980

tttcccgccc ccgccaaaac cgctgccacg tcgtcgtctc ccgccaaaaa aaaaaaagta 2040

ggctttggga tcgccgccgg tgacgatgaa gcca 2074

<210> 2

<211> 41

<212> DNA

<213>artificial sequence (Artificial Sequence)

<400> 2

cgacggccag tgccacgttg tcaggcctac tgggtaagtg c 41

<210> 3

<211> 44

<212> DNA

<213>artificial sequence (Artificial Sequence)

<400> 3

gaaatttacc ctcagatcta ccattggctt catcgtcacc ggcg 44

<210> 4

<211> 27

<212> DNA

<213>artificial sequence (Artificial Sequence)

<400> 4

gatcagttta aagaaagatc aaagctc 27

<210> 5

<211> 25

<212> DNA

<213>artificial sequence (Artificial Sequence)

<400> 5

ctgcaaggcg attaagttgg gtaac 25

<210> 6

<211> 21

<212> DNA

<213>artificial sequence (Artificial Sequence)

<400> 6

cttagccaga cgagcgggtt c 21

<210> 7

<211> 19

<212> DNA

<213>artificial sequence (Artificial Sequence)

<400> 7

gcttctgcgg gcgatttgt 19

Claims (10)

1. Rice Anther and pollen specific promoter PCHF17, which is characterized in that it is with following any nucleotides sequence Column:

1) nucleotide sequence shown in SEQ ID NO.1；

2) one or more nucleotide are substituted, lack or added in the nucleotide sequence shown in SEQ ID NO.1 and are had The function of same anther and pollen specific promoter, the nucleotide sequence as derived from 1)；

3) nucleotide sequence complementary with nucleotide sequence shown in SEQ ID NO.1.

2. the expression casette containing Rice Anther described in claim 1 Yu pollen specific promoter PCHF17.

3. the expression vector containing Rice Anther described in claim 1 Yu pollen specific promoter PCHF17.

4. the host cell containing expression vector described in expression cassette described in claim 2 or claim 3.

5. Rice Anther described in claim 1 and pollen specific promoter PCHF17 or the expression cassette containing it, expression carry Body or host cell are in the application for driving foreign gene specific expressed in plant anther and pollen.

6. Rice Anther described in claim 1 and pollen specific promoter PCHF17 or the expression cassette containing it, expression carry The application of body or host cell in prepare transgenosis plant.

7. Rice Anther described in claim 1 and pollen specific promoter PCHF17 or the expression cassette containing it, expression carry The application of body or host cell in prepare transgenosis rice.

8. expanding the primer pair of Rice Anther and pollen specific promoter PCHF17 described in claim 1, which is characterized in that institute The nucleotides sequence for stating primer pair is classified as nucleotide sequence shown in SEQ ID NO:2-3.

9. Rice Anther described in claim 1 and pollen specific promoter PCHF17 or the expression cassette containing it, expression carry Body or host cell or primer pair according to any one of claims 8 are preparing foreign gene in turn of Rice Anther and pollen-specific expression Application in trans-genetic hybrid rice.

10. the method for driving foreign gene specific expressed in plant anther and pollen, which is characterized in that including walking as follows It is rapid: Rice Anther and pollen specific promoter PCHF17 and purpose foreign gene are imported into carrier and obtained containing PCHF17 With the recombinant expression carrier of the expression cassette of purpose foreign gene, and it is introduced into Plant Genome, obtains foreign gene in anther With genetically modified plants specifically expressed in pollen.