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WO2012031421A1 - Gène du variant de l'epsp synthétase du coton et ses utilisations - Google Patents

Gène du variant de l'epsp synthétase du coton et ses utilisations Download PDF

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WO2012031421A1
WO2012031421A1 PCT/CN2010/078327 CN2010078327W WO2012031421A1 WO 2012031421 A1 WO2012031421 A1 WO 2012031421A1 CN 2010078327 W CN2010078327 W CN 2010078327W WO 2012031421 A1 WO2012031421 A1 WO 2012031421A1
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gene
epsps
glyphosate
cotton
mutant
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Chinese (zh)
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王建胜
何云蔚
崔洪志
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BIOCENTURY TRANSGENE(CHINA) Co Ltd
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BIOCENTURY TRANSGENE(CHINA) Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1085—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)
    • C12N9/1092—3-Phosphoshikimate 1-carboxyvinyltransferase (2.5.1.19), i.e. 5-enolpyruvylshikimate-3-phosphate synthase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8274—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for herbicide resistance
    • C12N15/8275—Glyphosate

Definitions

  • the invention belongs to the field of plant genetic engineering research, and particularly relates to cotton EPSP Modification of synthetase gene and construction of expression vector, and application in the development of glyphosate-tolerant transgenic plants.
  • Glyphosate is a non-selective herbicide with the advantages of broad-spectrum weeding and rapid degradation in soil. It is one of the safest herbicides for humans and animals because it is not present in animals. Glyphosate has the advantages of stable physical and chemical properties, high efficiency, broad spectrum, low toxicity, low residue, easy to be decomposed by microorganisms, and does not damage the soil environment. It has been widely used in agricultural production and has become the most abundant pesticide variety in the world. since Glyphosate herbicide, Monsanto, USA, 1976 - Roundup Since the successful development and wide application, the glyphosate-to-glycan transgenic research has become a hot spot in the research of herbicide resistance genetic engineering. With the development of glyphosate-resistant gene clones, glyphosate-tolerant GM crops have also been introduced and widely applied.
  • the mechanism of action of glyphosate is mainly competitive inhibition of the shikimate pathway 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) Activity.
  • EPSPS 5-enolpyruvylshikimate-3-phosphate synthase
  • the enzyme is a key enzyme in the biosynthesis of aromatic amino acids (including tryptophan, tyrosine, phenylalanine) in fungi, bacteria, algae and higher plants.
  • Glyphosate is a dilute phosphate pyruvate
  • PEP a competitive inhibitor of EPSPS, which combines glyphosate, EPSPS, and shikimic acid trisphosphate (S3P) to form an EPSPS-S3P-glyphosate complex ( This complex is very stable), suppress EPSPS
  • S3P shikimic acid trisphosphate
  • the activity of the branched acid is blocked, blocking the biosynthesis of aromatic amino acids and some aromatic compounds, which eventually leads to the metabolic imbalance of some hormones and key metabolites such as flavonoids, lignin and phenolic compounds, thus disturbing the normality of the organism. Nitrogen metabolism causes it to die.
  • the selection of the resistant EPSPS gene is a key factor in obtaining transgenic crops.
  • Change EPSPS The amino acid sequence of the functional region reduces its affinity for glyphosate while maintaining the catalytic activity of the enzyme. Stalker DM, etc. (1985 The chemical mutagen was used to treat Salmonella typhimurium, and the aroA gene mutant was cloned from the glyphosate resistant mutant. The 101st Ser in the mutant was confirmed by DNA and protein sequence analysis. The pro-gene was transferred into E. coli to obtain glyphosate-resistant properties. Sost D et al. (1990) cloned and sequenced wild-type Klebsiella.
  • AroA of pneumoniae and glyphosate-resistant mutant Klebsiella pneumoniae K1 (which encodes a glyphosate-insensitive EPSPS)
  • the gene which shows a single base difference, causes the 96-position Gly to become Ala in the amino acid sequence.
  • Baerson SR et al found a glyphosate-resistant goosegrass in the Malay Islands (Eleusine indica), which has a 2-4 fold higher LD50 than other glyphosate-sensitive species in the area, by comparing the EPSPS of resistant Goosegrass and Sensual Goosegrass The sequence was found to have two amino acids, one of which was an intentional mutation, ie the 106th Pro of the resistant Goosegrass EPSPS was replaced by Ser.
  • EPSPS Only transport to the chloroplast can play a role. Therefore, when constructing a plant expression vector, a peptide sequence needs to be added before the gene to form a fusion expression, which facilitates the targeted transfer of the target protein into the chloroplast.
  • the present invention selects cotton
  • the chloroplast-derived peptide (CTP) of the EPSPS gene serves as a fusion expression leader peptide of the gene of interest.
  • the object of the present invention is to provide a cotton 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS)
  • EPSPS cotton 5-enolpyruvylshikimate-3-phosphate synthase
  • a second object of the present invention is to provide an amino acid sequence of a cotton EPSPS mutant, such as SEQ ID NO: 2 Shown.
  • a third object of the present invention is to provide a nucleotide sequence of a cotton EPSPS mutant fusion gene, such as SEQ ID NO : 3 is shown.
  • a fourth object of the present invention is to provide an amino acid sequence of a cotton EPSPS mutant fusion gene, such as SEQ ID NO : 4 is shown.
  • a fifth object of the present invention is to provide a plant expression vector comprising the cotton EPSPS mutant gene or a fusion gene thereof.
  • a sixth object of the present invention is to provide a plant cell, tissue or plant transformed with the plant expression vector.
  • a seventh object of the present invention is to use the cotton EPSPS mutant gene or a fusion gene thereof in a glyphosate resistant plant variety.
  • the primers of NCPS were used to design primers to amplify cotton EPSPS and its fusion peptide sequences, which were named C- EPSPS , CTP.
  • the C- EPSPS gene was modified by multi-point mutation technology to obtain the mutant gene MC- EPSPS .
  • prokaryotic expression product glyphosate-resistant function construct prokaryotic expression vector of MC- EPSPS gene, transform expression plasmid BL21 (DE3) PlySs , culture under selective pressure of glyphosate, observe its growth, and turn The expression bacteria of C -EPSPS and empty vector were used as controls. The expression of MC -EPSPS and C- EPSPS under normal induction conditions was analyzed.
  • a plant expression vector containing MC -EPSPS the method comprising: selecting a 35S promoter containing a double enhancer and Tnos as a promoter and terminator of the MC- EPSPS gene, and adding an OK sequence after the promoter, The stop codon of the gene is followed by a PS sequence.
  • the chloroplast-derived peptide sequence (CTP) of the cotton EPSPS gene was added to the 5' end of the gene of interest to obtain fusion expression in plants.
  • the entire expression cassette was inserted into the modified binary expression vector pBI121 to obtain a glyphosate resistant plant expression vector pBI-MC- EPSPS . Transformation of tobacco and cotton by Agrobacterium-mediated transformation.
  • Figure 1 Growth curves of transformants containing MC- EPSPS- pET30a, C- EPSPS- pET30a and pET30a in M9 medium containing 100 mM glyphosate
  • Figure 2 Growth curve of BL21 (DE3) PlysS transformants containing MC- EPSPS- pET30a, C- EPSPS- pET30a and pET30a in M9 liquid medium containing 150 mM glyphosate
  • FIG 3 A growth is EPSPS deficient strain ER2799 containing plasmid MC -EPSPS -pET30a on M9 solid medium; B is EPSPS deficient strain ER2799 containing plasmid C -EPSPS -pET30a on M9 solid medium Growth condition; C is the growth of EPSPS-deficient strain ER2799 containing pET30a plasmid in M9 solid medium
  • Figure 4 Protein expression of BL21 (DE3) PlysS transformants containing MC- EPSPS -pET30a, C- EPSPS -pET30a and pET30a, respectively
  • Figure 5 Roadmap for constructing plant expression vector pBI-MC- EPSPS
  • Figure 6 A is the result of tobacco application to the MC-EPSPS gene after 0.2% glyphosate 7 days; B Non-GMO tobacco smeared 0.2% glyphosate results after 7 days
  • Figure 7 A is the result of spraying 0.2% glyphosate on cotton for MC-EPSPS gene for 7 days; B Is the result of non-GM cotton spraying 0.2% glyphosate after 7 days
  • EPSPS gene of cotton was named C- EPSPS .
  • the amplification primer sequence is as in Appendix SEQ ID No: 5, SEQ ID No: 6:
  • PCR conditions 94 °C 5min, 94 °C 45s, 56 °C 45s, 72 °C 4min , 5 cycles; 94 °C 45s, 60 °C 45s, 72 °C 4min, 25 cycles; 72 °C 7min.
  • the amplified product was digested with EcoR I and Sac I and constructed into the cloning vector pBulescript.
  • the vector was named pBulescript-C- EPSPS .
  • Phosphorylation of the 5' end of the primer facilitates the ligation and cyclization of subsequent PCR products.
  • the phosphorylation process is as follows:
  • the multi-point mutation was carried out using the cloning vector pBulescript-C- EPSPS containing the cotton EPSP synthase gene as a template, and the reaction procedure was carried out in accordance with the instructions in the QuikChange Multi Mutation Kit of STRATAGENE.
  • the vector after the mutation was named pBulescript-MC- EPSPS .
  • the mutant nucleotide sequence is as in Appendix SEQ ID No: 1, and the amino acid sequence is as in Appendix SEQ ID No: 2
  • the nucleotide homology with the original gene is 99.55%, and only the amino acids at the 102nd and 106th positions are changed at the amino acid level, and the homology is also 99.55%. Mutant clones by PCR And after sequencing and correct identification, save and reserve.
  • the primers were designed to amplify the MC-EPSPS gene, and the Nde I restriction site was added to the 5' end, and the Sac I restriction site was added to the 3' end.
  • the mutated pBulescript-MC- EPSPS was used as a template, and the primer sequence is shown in Appendix SEQ ID. No: 10, SEQ ID No: 11.
  • PCR conditions 94 ° C 5 min, 94 ° C 45 s, 54 ° C 45 s, 72 ° C 4 min, 30 cycles; 72 ° C 7 min.
  • the PCR product was digested with Nde I and Sac I and constructed into prokaryotic expression vector pET30a.
  • the recombinant expression vector MC- EPSPS- pET30a was obtained and transformed into prokaryotic expression strain BL21 (DE3) PlysS .
  • Invertants BL21 (MC- EPSPS -pET30a), BL21 (C- EPSPS -pET30a) and BL21 (pET30a) were inoculated into liquid M9 basal medium (including Cannamycin) 50 ⁇ g/mL), after activation at 37 °C overnight, dilute 1:100, inoculate 300 ⁇ L into 30 mL liquid M9 basal medium (containing kanamycin 50 ⁇ g/mL) at 200 rpm , 37 ° C air bath conditions.
  • liquid M9 basal medium including Cannamycin 50 ⁇ g/mL
  • Lanes 7 and 8 are BL21 (pET30a) crude extracts of two clone proteins. It can be seen from the electropherogram that the IPTG-inducible clone carrying the EPSPS gene can express the target protein of about 47 kDa in size. It was shown that both wild-type and mutant genes were efficiently expressed under the same expression conditions, and there was no significant difference in expression levels.
  • MC-EPSPS The increase in resistance is not due to overexpression of the gene, but is due to changes in protein structure.
  • transformants BL21 (MC -EPSPS -pET30a), BL21 (C- EPSPS -pET30a) and BL21 (pET30a) were inoculated into each inoculated into a liquid LB medium, 37 [deg.] C, Incubate for 12 h at 200 rpm.
  • the plasmid was extracted by alkaline lysis method, and 0.1 ⁇ g of the plasmid was transformed into EPSP synthase-deficient strain Escherichia coli ER2799 .
  • the transformants were cultured on M9 solid medium containing 1 mM IPTG and kanamycin 50 ug/ml for 36 h.
  • ER2799 MC- EPSPS -pET30a
  • C- EPSPS -pET30a ER2799
  • No transformants appeared. It is indicated that the mutant protein still has certain enzymatic activity.
  • the OK- Pst I- Xho I - PS fragment was synthesized according to the published OK (Omega & Kozak) sequence and the PS (Processing & Splicing sequence) sequence (both fully disclosed in ZL 95 119563.8) with Bam HI and The Sac I restriction site is ligated between the OK and PS by Pst I and Xho I restriction sites, and the protection bases are inserted between the two restriction sites to facilitate subsequent restriction enzyme construction, using Bam HI and Sac I.
  • the OK- Pst I- Xho I - PS was cloned into the recombinant plasmid 35S-pUC18 to obtain the recombinant plasmid 35S-OK-PS-pUC18.
  • the CTP - MC- EPSPS fragment was amplified by using pBulescript-MC- EPSPS as a template.
  • the gene sequence is shown in SEQ ID No: 3, and the Pst I and Xho I restriction sites were added to the ends of the fusion sequence, and the primer sequence was SEQ ID.
  • CTP-MC- EPSPS was ligated into 35S-OK-PS-pUC18 using the restriction sites at both ends to obtain recombinant plasmid 35S-OK-CTP-MC-PS-pUC18. .
  • the 35S-PS fragment of 35S-OK-CTP-MC-PS-pUC18 was cloned into pBI121 by using Hin d III and Sac I, and the original 35S - GUS was replaced to obtain the recombinant plant expression vector pBI-MC- EPSPS .
  • the process is shown in Figure 5 of the accompanying drawings.
  • the obtained transgenic tobacco leaves were extracted and identified by PCR.
  • the tobacco leaves which were positive by 0.2% glyphosate spray PCR were observed. After 7 days, the experimental results were observed.
  • the MC was transferred to MC- The EPSPS gene transgenic tobacco grew normally (Fig. 6A), while the non-transgenic tobacco growth was significantly inhibited, and the leaves were yellow and wilting (Fig. 6B).
  • Agrobacterium-mediated transformation is a plant genetic transformation method well known to researchers in the field. The specific operating procedures are:
  • the constructed pBI-MC- EPSPS plasmid was electroporated into Agrobacterium strain LBA4404 , and Agrobacterium single colony was inoculated into LB or YEB liquid medium containing kanamycin 50 mg/L and rifampicin 25 mg/L. in.
  • the dark culture was shaken overnight at 28 ° C until the logarithmic phase of bacterial growth. Dilute the bacterial solution with LB or YEB liquid medium, shake it for 4-6 hours, and dilute the bacterial solution to an OD600 value of 0.3 to 0.35.
  • the cotton seeds are desulfurized with sulfuric acid (H 2 SO 4 ), the sulfuric acid on the surface of the seeds is washed away with tap water, dried, and the seeds are surface-sterilized with 70% ethanol for 1 min, and then peroxidized with 10% to 15%.
  • the co-cultured hypocotyl segments were placed in callus induction medium (MS + 2,4-D 0.1 mg/L + KT 0.1 mg/L + MgCl 2 0.91 g/L + Gelrite 2.0 g/L + Kanamycin 50 ⁇ 100 mg / L + cephalosporin 500 mg / L + glucose 30 g / L, pH 5.8), cultured under normal conditions (25 ° C) for 2 months (change the same medium once a month) ).
  • the callus was picked a little and the selection marker gene nptII was detected.
  • the callus with positive test results continued to be subcultured, and the non-positive callus was eliminated.
  • the frequency of cotton resistant callus was 50% ⁇ 76%.
  • the induced resistant callus was introduced into a proliferation medium (MS medium + MgCl 2 0.91 g/L + Gelrite 2.0 g/L + glucose 30 g/L, pH 5.8), and cultured under normal conditions (25 ° C). , once every other month, until the callus differentiates. After the first and second transfer to the proliferation medium, some of the callus browning died, and the normal callus did not proliferate rapidly. After the second passage, the callus proliferation rate was accelerated.
  • a proliferation medium MS medium + MgCl 2 0.91 g/L + Gelrite 2.0 g/L + glucose 30 g/L, pH 5.8
  • the planted regenerated cotton seedlings are placed in an artificial incubator with a temperature control of 22 ° C and a humidity control of 80 to 85% for 5 to 7 days, and then cultured in a greenhouse for 10 to 20 days, and then transplanted into a soil pot or a field.
  • the herbicide-tolerant gene plant expression vector was introduced into cotton to obtain transgenic cotton.
  • the identification method was as in Example 6.
  • the results were as shown in Figures 7A and 7B in the accompanying drawings.
  • the transgenic cotton transgenic with MC- EPSPS gene was able to grow normally (Fig. 7A), while the growth of non-transgenic tobacco was significantly inhibited, and the leaves were yellow and wilting. (Fig. 7B).

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Abstract

Cette invention concerne un variant de l'EPSP synthétase du coton et un gène mutant artificiel MC-EPSPS codant pour ledit variant de l'EPSP synthétase. Le gène mutant selon l'invention codant pour le variant contenant deux sites d'acides aminés mutants et ayant une propriété anti-glyphosate est obtenu par conception des sites mutants selon des documents et l'analyse de la structure protéique, puis mise en œuvre d'une amplification par PCR à l'aide du gène de l'EPSP synthétase de Gossypium hirsutum à titre de matrice, avec recombinaison homologue du gène et mutations multisites de la séquence d'ADN. La performance de liaison de l'EPSP synthétase codée par le gène mutant au glyphosate est réduite, de manière dominante, et l'EPSP synthétase obtenue possède une certaine efficacité de catalyse enzymatique. Le gène codant pour le variant de l'EPSP synthétase selon l'invention peut être utilisé pour cultiver de nouvelles espèces de plantes transgéniques ayant une propriété anti-glyphosate.
PCT/CN2010/078327 2010-09-08 2010-11-02 Gène du variant de l'epsp synthétase du coton et ses utilisations Ceased WO2012031421A1 (fr)

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CN2010102776678A CN102399794A (zh) 2010-09-08 2010-09-08 一种棉花epsp合成酶突变体基因及其应用
CN201010277667.8 2010-09-08

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103205404A (zh) * 2013-03-26 2013-07-17 上海市农业科学院 来源于苹果的epsp合酶多位点突变体及其编码基因与应用
CN111394368A (zh) * 2020-04-29 2020-07-10 海南大学 第182位点发生突变的橡胶树epsps基因及其应用

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* Cited by examiner, † Cited by third party
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CN102776158B (zh) * 2012-06-14 2013-10-23 重庆市农业科学院 一种抗草甘膦EPSP合成酶GmEPSPS-2及其编码基因与应用
CN102816777A (zh) * 2012-09-06 2012-12-12 南京农业大学 植物抗/耐草甘膦基因及其应用
CN102876690A (zh) * 2012-10-09 2013-01-16 上海市农业科学院 来源于肺炎克雷伯氏杆菌342的epsp合酶基因及其应用
WO2015027369A1 (fr) * 2013-08-26 2015-03-05 创世纪种业有限公司 Protéine de fusion anti-glyphosate et gène codant pour elle, procédé de préparation et utilisation
CN109022385B (zh) 2017-12-25 2020-02-14 四川天豫兴禾生物科技有限公司 一种含l195p和s247g突变的植物epsps突变体及其编码基因和应用
CN110229843B (zh) * 2019-04-04 2023-02-28 中国农业科学院棉花研究所 陆地棉转化事件19pfa1-135-17及其特异性鉴定方法
CN111139254A (zh) * 2020-01-15 2020-05-12 吉林农业大学 大豆GmEPSPS1和GmEPSPS2定向突变修饰基因及其克隆方法和应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1196088A (zh) * 1995-07-19 1998-10-14 罗纳-普朗克农业公司 突变的5-烯醇丙酮酰莽草酸－3－磷酸合酶,编码此蛋白的基因以及含有此基因的转化的植物
US6040497A (en) * 1997-04-03 2000-03-21 Dekalb Genetics Corporation Glyphosate resistant maize lines
CN101619319A (zh) * 2009-03-13 2010-01-06 创世纪转基因技术有限公司 人工改造合成的抗草甘膦基因与应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007146980A2 (fr) * 2006-06-13 2007-12-21 Athenix Corporation Epsp synthases améliorées, compositions et procédés d'utilisation

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1196088A (zh) * 1995-07-19 1998-10-14 罗纳-普朗克农业公司 突变的5-烯醇丙酮酰莽草酸－3－磷酸合酶,编码此蛋白的基因以及含有此基因的转化的植物
US6040497A (en) * 1997-04-03 2000-03-21 Dekalb Genetics Corporation Glyphosate resistant maize lines
CN101619319A (zh) * 2009-03-13 2010-01-06 创世纪转基因技术有限公司 人工改造合成的抗草甘膦基因与应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DATABASE GENBANK 17 September 2007 (2007-09-17), Database accession no. ABV24481.1 *
DATABASE GENBANK 17 September 2007 (2007-09-17), Database accession no. EU123855.1 *
TONG XUHONG ET AL.: "Cloning of 5-enolpyruvylshikimate-3-phosphate Synthase Genes EPSPS from the Upland Cotton and Its Expression in Different Cotton Tissues", COTTON SCIENCE, vol. 21, no. 4, 31 August 2009 (2009-08-31), pages 263 - 264 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103205404A (zh) * 2013-03-26 2013-07-17 上海市农业科学院 来源于苹果的epsp合酶多位点突变体及其编码基因与应用
CN103205404B (zh) * 2013-03-26 2014-08-27 上海市农业科学院 来源于苹果的epsp合酶多位点突变体及其编码基因与应用
CN111394368A (zh) * 2020-04-29 2020-07-10 海南大学 第182位点发生突变的橡胶树epsps基因及其应用
CN111394368B (zh) * 2020-04-29 2023-04-18 海南大学 第182位点发生突变的橡胶树epsps基因及其应用

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