CN102559685B - Anther tapetum specific expression promoter POsABCG15, recombinant expression vector thereof and application thereof - Google Patents
Anther tapetum specific expression promoter POsABCG15, recombinant expression vector thereof and application thereof Download PDFInfo
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Abstract
The invention discloses an anther tapetum specific expression promoter POsABCG15. The promoter has a nucleotide sequence shown as SEQ ID NO. 31. The promoter can regulate and control a target gene, so that the target gene is expressed to anther tapetum specifically; and the POsABCG15 in rice regulates and the controls the expression of a lipid deposition gene OsABCG15 on anther epidermis and pollen wall of the rice and influences the accumulation of lipids on surfaces of anther and pollen, so that the anther tapetum specific expression promoter POsABCG15 can be used for generating a new rice male sterile line or regulating and controlling the expression of other genes in the tapetum, and has an important effect in agricultural production.
Description
Technical field
The invention belongs to the genetically engineered field, particularly anther tapetum specific expression promoter
POsABCG15, also relate to anther tapetum specific expression promoter
POsABCG15Recombinant expression vector and application.
Background technology
Paddy rice (
Oryza sativa) be one of staple food crop, there is half population in the whole world approximately take rice as staple food.In China, the rice growing area is 30% of national food crop sown area, and ultimate production accounts for more than 42% of food crop output, and its major reason is exactly the successful Application of rice heterosis.Jones has just found the rice heterosis phenomenon as far back as nineteen twenty-six.But paddy rice is the self-fertility crop, is difficult to obtain cross-fertilize seed a large amount of, that purity is high, has hindered the process of rice heterosis utilization, until China Lee in 1970 must lose sterile strain in discovery open country, lake." father of hybrid rice " Yuan Longping's in 1973 research team through diligently specially grinding, has realized three series mating, and beginning spread hybrid rice in 1976 becomes a reality rice heterosis.Up to now, sterile line is still the main path of rice heterosis utilization.
At present, people mainly realize the hybrid vigour of paddy rice by Cytoplasm nuclear interaction male sterile line (three series) and warm photosensitive sterile line (bilinear method).Yet, because the Cytoplasm nuclear interaction male sterile line combo is not freely, available parent is limited, seed is produced more complicated than bilinear method, the fertility of temperature photosensitive sterile line is affected by nature temperature optical condition, the reasons such as the big area risk in hybrid seed production is large are so that the hybrid vigour of paddy rice can't find full expression by these two kinds of sterile lines.Anther tapetum specific expression promoter can be regulated and control the downstream gene specifically expressing and organizes in anther tapetum, and obtains male sterile line by the growth that anther tapetum specifically expressing genes involved can affect flower pesticide and pollen.Therefore, be badly in need of a kind of anther tapetum specific expression promoter, can provide strong instrument for utilizing genetic engineering means acquisition male sterile line and restorer.
Summary of the invention
In view of this, one of purpose of the present invention is to provide anther tapetum specific expression promoter
POsABCG15, this promoter regulation goal gene specifically expressing is in anther tapetum, and the control microspore development becomes the expression of the genes involved of fertile pollen, can be used in and produces new male sterible series of rice, plays an important role in agriculture production, and technical scheme is:
Anther tapetum specific expression promoter
POsABCG15, described anther tapetum specific expression promoter
POsABCG15Nucleotide sequence shown in SEQ ID NO.31.
Two of purpose of the present invention is to provide anther tapetum specific expression promoter
POsABCG15Recombinant expression vector, its technical scheme is:
Contain described anther tapetum specific expression promoter
POsABCG15Recombinant expression vector.
Preferably, described recombinant expression vector is at the pCAMBIA130021 carrier
GUSBe written into anther tapetum specific expression promoter before the gene
POsABCG15
Three of purpose of the present invention is to provide anther tapetum specific expression promoter
POsABCG15Application, technical scheme is:
Described anther tapetum specific expression promoter
POsABCG15In the application of mediate foreign gene specifically expressing in anther tapetum.
Further, described foreign gene is
OsABCG15Gene.
Beneficial effect of the present invention is: anther tapetum specific expression promoter disclosed by the invention
POsABCG15, this promoter regulation goal gene specifically expressing is in anther tapetum, adjusting and controlling rice flower pesticide epidermis and pollen wall lipid deposition gene in paddy rice
OsABCG15Express, affect flower pesticide and pollen surface lipid accumulation, being obstructed when expression to produce male sterible series of rice; The invention also discloses anther tapetum specific expression promoter
POsABCG15Recombinant expression vector, can be as the promotor of anther tapetum specific expression gene, at the anther tapetum specific expression promoter of recombinant expression vector
POsABCG15Make downstream gene be positioned anther tapetum under the regulation and control and express, for plant genetic engineering field research gene provides instrument at the anther tapetum specifically expressing, be with a wide range of applications.
Description of drawings
In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention is described in further detail below in conjunction with accompanying drawing, wherein:
Figure l is the present invention
Osabcg15Mutant and normal near isogenic line II-32B morphology of the anther and pollen are learned and are observed synoptic diagram (Sp represents the pollen that dyes, and Ac represents the flower pesticide residue).
Fig. 2 is the present invention
OsABCG15Location, seat and mutational site synoptic diagram.
Fig. 3 is anther tapetum specific expression promoter of the present invention
POsABCG15Regulation and control
OsABCG15Gene specific expression analysis figure.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.These embodiment only are not used in for explanation the present invention and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, Sambrook equimolecular cloning experimentation chamber handbook (New York:Cold Spring Harbor Laboratory Press for example, 1989) condition described in, or the condition of advising according to manufacturer.
Term " gene of flower pesticide epidermis and pollen wall lipid deposition " refers to encode and has transportation lipid to the nucleotide sequence of the ABC-2 type translocator of flower pesticide epidermis and pollen wall, nucleotide sequence and the degenerate sequence thereof of base sequence shown in 1-2067 position among the SEQ ID NO.2.Degenerate sequence refers to, is arranged in the encoder block 1-2067 position Nucleotide of SEQ ID NO. 2 sequences, the sequence that has one or more codons to be encoded to produce after the degenerate codon of same amino acid replaces.Because the degeneracy of codon, thus with SEQ ID NO.2 in 1-2067 position nucleotide sequence homology be low to moderate about 70% the degenerate sequence described sequence of SEQ ID NO.1 of also encoding out.This term also comprises can be under the rigorous condition of moderate, better under highly rigorous condition with SEQ ID NO.2 in from the nucleotide sequence of the nucleotide sequence hybridization of the 1 one 2067 in Nucleotide.This term also comprise with SEQ ID NO.2 in from the homology at least 70% of the nucleotide sequence of the 1 one 2067 in Nucleotide, preferably at least 80%, more preferably at least 90%, at least 95% nucleotide sequence best.This term also comprises encoding to have the variant form of open reading frame sequence among the SEQ ID NO.2 with the albumen of the identical function of natural regulation and control flower pesticide and pollen wall lipid accumulation.These variant forms comprise (but being not limited to): several (are generally 1-90, preferably 1-60, more preferably 1-20,1-10 best) disappearance, insertion and/or the replacement of Nucleotide, and several (are generally in 60 to hold interpolation 5 ' and/or 3 ', preferably being in 30, more preferably is in 10, is in 5 best) Nucleotide.
Term " albumen of flower pesticide epidermis and pollen wall lipid deposition " refers to have transportation lipid to polypeptide ABC-2 type translocator activity, that have SEQ ID NO.1 sequence of flower pesticide epidermis and pollen wall.This term also comprises having and the natural variant form that affects the ABC-2 type translocator SEQ ID NO.1 sequence of flower pesticide epidermis and pollen wall lipid deposition.These variant forms comprise (but being not limited to): several (are generally 1-50, preferably 1-30, more preferably 1-20,1-10 best) amino acid whose disappearance, insertion and/or replacement, and add one or several at C-terminal and/or N-terminal and (be generally in 20, preferably being in 10, more preferably is in 5) amino acid.In the art, when replacing with the close or similar amino acid of performance, usually can not change the function of protein; Add one or the common function that also can not change protein of several amino acid at C-terminal and/or N-terminal.
Term " promotor " refers to the section of DNA sequence for RNA polymerase identification, combination and transcriptional start.Synthetic being called of RNA, transcribe under DNA instructs, the initial promotor control by DNA of transcribing.
Among the embodiment, can select various carrier known in the art, the carrier as commercially available comprises plasmid, clay etc.When the Rice Anther epidermis of producing present embodiment and pollen wall lipid transhipment related polypeptide, the encoding sequence of this albumen operationally can be connected in expression regulation sequence, thereby form the expression vector of adjusting and controlling rice flower pesticide epidermis and pollen wall lipid transfer related protein.
Associated nucleotide full length sequence or its fragment can obtain with the method for pcr amplification method, recombination method or synthetic usually.For the pcr amplification method, can be according to the disclosed relevant nucleotide sequence of present embodiment, especially open reading frame sequence designs primer, and with commercially available cDNA storehouse or by the prepared cDNA storehouse of ordinary method well known by persons skilled in the art as template, amplification and must relevant sequence.When sequence is longer, usually needs to carry out twice or pcr amplification repeatedly, and then the fragment that each time amplifies is stitched together by proper order.In case obtained relevant sequence, just can obtain in large quantity relevant sequence with recombination method.This normally is cloned into carrier with it, changes cell over to again, then separates obtaining relevant sequence from the host cell after the propagation by ordinary method.
Except producing with recombination method, the fragment of albumen available solid phase technique is also produced by direct peptide synthesis among the embodiment.Can carry out by hand or automatically at external synthetic protein.Can distinguish each fragment of chemosynthesis present embodiment albumen, then be connected to produce the molecule of total length with chemical process.
Extensive No. 1 mutant of long-grained nonglutinous rice red silk, called after
Osabcg15Mutant is preserved by this laboratory, makes recurrent parent with II-32B, with
Osabcg15Mutant hybridization and throwback backcross, and mutant preserves the most at last, and this sterile material is a pair of near isogenic line with II-32B.
Osabcg15Mutant and II-32B hybridization, Fl is for being to educate all, separating appears in generation in selfing F2, wherein normal plant is 567, mutant strain is 187, and ratio meets 3:1 (χ 2=0.0071＜χ 0.05=3.84), shows that this male-sterile mutation body surface type is caused by a monokaryon transgenation.Right
Osabcg15The morphological observation of mutant plant: II-32B spike of rice solid rear sagging (Figure 1A is left), and
Osabcg15The shaky vertical (Figure 1A is right) that is of mutant small ear; II-32B flower pesticide comprises mature pollen and is rendered as yellow (on Figure 1B),
Osabcg15Mutant flower pesticide is white (under Figure 1B); II-32B pollen iodine dyes and is black (on Fig. 1 C),
Osabcg15WUHUAFEN in the sudden change flower pesticide only has flower pesticide residue (under Fig. 1 C) after smashing to pieces in iodine liquid.
Embodiment 2,
OsABCG15The location of gene and clone
One, target group
Will
Osabcg15The fine hybridization of mutant and japonica rice Japan, selfing obtains F2 generation, and the selection male sterile plants is target group.
Two, extract paddy DNA
The parent adopts modified CTAB method to extract, comprising the steps: to get blade 0.1-0.2 gram (about half sheet) is put in the little mortar, add an amount of liquid nitrogen, be ground at once powdery, the 2m1 centrifuge tube of packing into, add 1.5 * CTAB solution of 100 ℃ of preheatings of 700 μ L in centrifuge tube, put into 65 ℃ of water-baths behind the careful mixing, take out centrifuge tube after 20 minutes, add equal-volume chloroform/primary isoamyl alcohol, fierce mixing, centrifugal 10 minutes of 13000rpm, get supernatant in new pipe, add behind the 900 μ L dehydrated alcohol mixings-20 ℃ and put more than half an hour.The DNA that separates out is centrifugal, centrifugal 10 minutes of 14000rpm.Remove supernatant, will precipitate with the 1mL volume fraction is that 70% ethanol cleans once, and centrifugal drying is dissolved in the 200 μ L TE solution 4 ℃ of Refrigerator stores.The individual plant DNA of target group adopts improved alkaline-heating method to extract, and comprises the steps: to shred leaflet tablet 1-2cm
2Put into the 0.5m1 centrifuge tube, adding 100 μ L concentration is the NaOH solution of 0.125M, boiling water bath 30 seconds, then adding 50 μ L concentration is the Tris-HCl(pH8.0 of 1.0M), adding at last 100 μ L concentration is the HCl of 0.125M, boiling water bath after 2 minutes 4 ℃ of Refrigerator stores for subsequent use.
Three, colony's compartment analysis is just located
Design 137 pairs of polymorphism marks, comprising 42 pairs of SSR primers and 95 pairs of InDel molecule marker primers.Wherein the SSR primer is according to the sequence of having delivered synthetic (specifically referring to http://www.gramene.org.microsat/ssr.html), other InDel molecule marker designs are according to the nucleotide sequence of having announced that compares 9311 liang of strains of the japonica rice warm and fine long-grained nonglutinous rice of Japan, to the partial design primer of difference, verify 2 warm and fine long-grained nonglutinous rices of parent japonica rice Japan
Osabcg15Polymorphism between the mutant.137 pairs of primers of design are distinguished amplifying rice male sterile plants and parental rice genomic dna, the pcr amplification program is: in the 10 μ L systems, and l μ L template, l μ L 10pmol/ μ L upstream primer, l μ L10pmol/ μ L downstream primer, l μ L 10 * Buffer (Mg
20), l μ L 2mM dNTP, 0. l μ L Taq, 3.9 μ L water; The PCR product is 10% PAGE gel electrophoresis with the quality volume fraction, and silver staining method detects.The result shows that 137 pairs of marks carry out amplified reaction, and Chr6Ind(-37-1) primer (as shown in table 1) there are differences for the selection male sterile plants parent and F2, shows the mark Chr6Ind(-37-1 on the karyomit(e) No. 6) with
OsABCG15Locus has obvious linkage relationship.
Four, Fine Mapping
With sterile individual plant to the mark Chr6Ind(-37-1 on No. 6 karyomit(e)) near variant SSR mark verify, find that the RM3628 primer has respectively 6 and 38 s' recon and recon mutually not to comprise this explanation to (as shown in table 1) and RM5371 primer to (as shown in table 1)
OsABCG15Locus should be between RM3628 and two molecule markers of RM5371.For right
OsABCG15The further Fine Mapping of gene, to expand nearly 2157 strains to for the mutant strain of location, 20 pairs of SSR primers between RM3628 and RM5371, have been designed again, 6 pairs of InDel primers and 23 pairs of STS primers, RM20356, RM20361, RM20366, RM275 and RM275 primer are to there are differences in 20 pairs of SSR primers, Chr6Ind (30), Chr6Ind (7) and Chr6Ind (4) primer are to there are differences in 6 pairs of InDel primers, and Chr6STS20, Chr6STS13 and Chr6STS15 primer are to there are differences in 23 pairs of STS primers.With discrepant primer target group is analyzed, the most at last
OsABCG15The assignment of genes gene mapping is at Chr6Ind(-30) and Chr6STS13 between, shown in Fig. 2 A.Show that by analysis this zone only comprises 1 encoding gene, this genes encoding ABC-2 type translocator, the aminoacid sequence of this protein of encoding are shown in SEQ ID NO.1, and nucleotide sequence is shown in SEQ ID NO.2, with the Arabidopis thaliana male sterility gene
WBC27Homology by twice order-checking to this gene, is found
OsABCG15The replacement (A becomes C) of a base occurs in mutant at the 4th exon place of this gene, shown in Fig. 2 B.The sequence of the used mark of the assignment of genes gene mapping is as shown in table 1.
Table 1 assignment of genes gene mapping molecule marker and primer sequence thereof
| The primer title | Upstream primer (5 ' → 3 ') | Downstream primer (5 ' → 3 ') |
| RM3628 | 5’-aatcatgcctagagcatcgg-3’（SEQ ID NO.3） | 5’-gttcaacatgggtgcagatg-3’（SEQ ID NO.4） |
| RM20356 | 5’-ttaccaggcttcctctcttgacc-3’（SEQ ID NO.5） | 5’-ccacgtcacccagaaactaatcc-3’（SEQ ID NO.6） |
| RM20361 | 5’-cttgaaatttgtgcggaggttgc-3’（SEQ ID NO.7） | 5’-gatgtcaccatcacggagaattagg-3’（SEQ ID NO.8） |
| Chr6Ind(-30) | 5’-gcttccataactacaaggc-3’（SEQ ID NO.9） | 5’-atctcgtataacaaactcacaa-3’（SEQ ID NO.10） |
| Chr6Ind(4) | 5’-cgatattactaccaggattt-3’（SEQ ID NO.11） | 5’-attgccaaccaactaact-3’（SEQ ID NO.12） |
| Chr6STS20 | 5’-aaaatggtgggtcatacgg-3’（SEQ ID NO.13） | 5’-gggtcctgggtagcgaaa-3’（SEQ ID NO.14） |
| Chr6STS15 | 5’-gctccgtgaccaagtatgt-3’（SEQ ID NO.15） | 5’-gaggaagaagaagagggtgt-3’（SEQ ID NO.16） |
| Chr6STS13 | 5’-cgatgagatggttgggagc-3’（SEQ ID NO.17） | 5’-tgcgggcaggagatttgg-3’（SEQ ID NO.18） |
| Chr6Ind(-7) | 5’-acacgaggcttctagtgat-3’（SEQ ID NO.19） | 5’-attgattgttcctaaccaa-3’（SEQ ID NO.20） |
| RM20366 | 5’-caggtaaagcgatgagcaattcg-3’（SEQ ID NO.21） | 5’-aaggagttggcaacagcgaagg-3’（SEQ ID NO.22） |
| RM275 | 5’-cctcaacatcctcacacacaagc-3’（SEQ ID NO.23） | 5’-gccaatcggatgtgatttatgc-3’（SEQ ID NO.24） |
| Chr6Ind(-37-1) | 5’-atgtcctaagggtctgt-3’（SEQ ID NO.25） | 5’-agtggctacatttagtttg-3’（SEQ ID NO.26） |
| RM5957 | 5’-actgctgcactgcacaagac-3’（SEQ ID NO.27） | 5’-agctagctaggcgtgagctg-3’（SEQ ID NO.28） |
| RM5371 | 5’-ggctagctttagctgcgttg-3’（SEQ ID NO.29） | 5’-acccagatcgaaacaactgc-3’（SEQ ID NO.30） |
Analyze to find, be positioned at Fine Mapping
OsABCG15Upstream region of gene 2034bp has the RNA polymerase recognition site, and nucleotide sequence is shown in SEQ ID NO.31, and the called after anther tapetum specific expression promoter is called for short
POsABCG15According to primers shown in SEQ ID NO.31, upstream primer is
POsABCG15F:5'-gac
AagcttCgaaatgctcggttatatg-3 ' (SEQ ID NO.32), underscore is Hind III restriction enzyme site; Downstream primer is
POsABCG15 R:5'-gc
TctagaCtcctccaagagacacag-3 ' (SEQ ID NO.33), underscore is Xba I restriction enzyme site, increases with PCR, the PCR reaction conditions is: 95 ℃ of denaturations 3 minutes; 94 ℃ of sex change 45 seconds, 61 ℃ of annealing 45 seconds, 72 ℃ were extended 2 minutes, 28 circulations, last 72 ℃ were extended 10 minutes.The PCR product identifies that through agarose gel electrophoresis recovery contains anther tapetum specific expression promoter
POsABCG15The fragment of 2051bp size, with Hind III and Xba I enzyme cut-flower medicine tapetum specific expression promoter simultaneously
POsABCG15Pcr amplified fragment and pCAMBIA130021 carrier also reclaim anther tapetum specific expression promoter
POsABCG15With the pCAMBIA130021 carrier framework, with the anther tapetum specific expression promoter that reclaims
POsABCG15Be connected with the pCAMBIA130021 carrier framework, make
POsABCG15Promotor is written into the pCAMBIA130021 carrier
GUSBefore the gene, must contain anther tapetum specific expression promoter
POsABCG15Recombinant expression vector, called after:
POsABCG15:GUSCarrier.Will
POsABCG15:GUSCarrier transforms the Agrobacterium LBA4404 competent cell, obtains to contain
POsABCG15:GUSThe LBA4404 of carrier, called after LBA4404:
POsABCG15:GUSWith LBA4404:
POsABCG15:GUSInfect rice callus, obtain transgenic paddy rice.Activation analysis by beta-galactosidase (GUS)
POsABCG15Expression pattern, get transgenic paddy rice root, stem, the Ye Hehua of different developmental phases, dye by the GUS staining fluid, tissue after the dyeing is processed with 75% ethanol decolorization, and it is fixing that the material of decolouring processing is put into the FAA stationary liquid, dehydration, the processing such as infiltration and paraffin embedding, observe by section (l0um is thick), the result shows that the GUS dye liquor is blueness and is organized as anther tapetum, has shown to separate being positioned at of obtaining
OsABCG15Upstream region of gene 2034bp Nucleotide has promoter activity, and can regulate and control the downstream gene specifically expressing in anther tapetum, is anther tapetum specific expression promoter therefore.
In situ hybridization detects
POsABCG15The downstream
OsABCG15The expression tissue of gene, in situ hybridization uses probe to synthesize: with rice cDNA as template, upstream primer is OsABCG15IF:5'-agtaatacgactcactatagggacggccacctgacctac-3 ' (SEQ ID NO.34), downstream primer is OsABCG15IR:5'-agatttaggtgacactatagaacagcacccaaaccaaca-3 ' (SEQ ID NO.35), carry out pcr amplification, carry out mark with digoxin during amplification, make the in situ hybridization probe.Luo Shi (Roche) company's digoxin Yeast Nucleic Acid labelling kit (DIG RNA Labeling Kit (SP6/T7)) and digoxin kit for detecting nucleic acid (DIG Nucleic Acid Detection Kit) are used in situ hybridization, concrete steps are carried out according to the test kit specification sheets, wherein in situ hybridization result as shown in Figure 3, the result shows
OsABCG15Genetic expression has proved to be positioned in anther tapetum equally
OsABCG15Upstream region of gene 2034bp Nucleotide has promoter activity, and can regulate and control the downstream
OsABCG15Gene specific is expressed in anther tapetum, is anther tapetum specific expression promoter.
The invention is not restricted to use the pCAMBIA130021 carrier, also can use other can accept carrier; Make up
POsABCG15:GUSCarrier can be used as the expression vector of other genes, and goal gene is connected to
POsABCG15Replace in the downstream
POsABCG15:GUSGus gene in the carrier can be in anther tapetum specifically expressing.
Explanation is at last, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although by invention has been described with reference to the preferred embodiments of the present invention, but those of ordinary skill in the art is to be understood that, can make various changes to it in the form and details, and not depart from the spirit and scope of the present invention that appended claims limits.
Claims (2)
1. anther tapetum specific expression promoter
POsABCG15, it is characterized in that: described anther tapetum specific expression promoter
POsABCG15Nucleotide sequence shown in SEQ ID NO.31.
2. contain the described anther tapetum specific expression promoter of claim 1
POsABCG15Recombinant expression vector.
3, the described anther tapetum specific expression promoter of claim 1
POsABCG15In the application of mediate foreign gene specifically expressing in anther tapetum.
4, application according to claim 3 is characterized in that: described foreign gene is
OsABCG15Gene.
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| ES2913136T3 (en) * | 2012-11-09 | 2022-05-31 | Shenzhen Inst Of Molecular Crop Design | Fertility gene and uses of it |
| CN103725677B (en) * | 2013-11-18 | 2016-06-08 | 北京大北农科技集团股份有限公司 | Tissue-specific promoter and purposes thereof |
| CN109055606A (en) * | 2018-10-31 | 2018-12-21 | 宁夏泰金种业股份有限公司 | A kind of bumpkin hybrid seed purity identification method based on SSR marker method |
| CN109337999A (en) * | 2018-10-31 | 2019-02-15 | 宁夏泰金种业股份有限公司 | A kind of identification method of the muskmelon hybrid seed purity based on SSR marker method |
| CN109234437A (en) * | 2018-10-31 | 2019-01-18 | 宁夏泰金种业股份有限公司 | A kind of watermelon hybrid seed Purity method based on SSR marker method |
| CN110982819B (en) * | 2019-12-31 | 2023-05-05 | 海南波莲水稻基因科技有限公司 | Promoter PCHF7 specifically expressed in rice anther and pollen and application thereof |
| CN112219714B (en) * | 2020-08-27 | 2022-04-29 | 云南大学 | Method for breeding sporophyte recessive male nuclear sterility |
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Application publication date: 20120711 Assignee: Biotechnology Center of Beijing Dabeinong Technology Group Co., Ltd. Assignor: Southwest University Contract record no.: 2014990000013 Denomination of invention: Anther tapetum specific expression promoter POsABCG15, recombinant expression vector thereof and application thereof Granted publication date: 20130123 License type: Common License Record date: 20140108 |
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