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Plant EPSPS mutant with A138T mutation, and coding gene and application of mutant

A technology for encoding genes and mutants, applied in the field of genetic engineering, can solve the problem of low acceptance of transgenic crops, and achieve the effects of maintaining the catalytic activity of biological enzymes, improving resistance, and having broad application prospects.

Active Publication Date: 2019-01-01
SICHUAN GEVOTO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the resulting genetically modified crops have a considerable area of ​​promotion, due to the wave of anti-genetics, the acceptance of genetically modified crops in the world is still low. Even in the Americas with the largest planting area of ​​genetically modified crops, genetically modified crops are mainly limited to corn. , soybeans, cotton and several other crops

Method used

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  • Plant EPSPS mutant with A138T mutation, and coding gene and application of mutant
  • Plant EPSPS mutant with A138T mutation, and coding gene and application of mutant
  • Plant EPSPS mutant with A138T mutation, and coding gene and application of mutant

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Experimental program
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Effect test

Embodiment 1

[0095] This embodiment provides a plant EPSPS mutant, which is derived from rice and is rice EPSPS mutant I, which is obtained after mutation from wild-type rice EPSPS (amino acid sequence shown in SEQ ID NO: 5), and its amino acid sequence is as follows: Shown in SEQ ID NO:6.

[0096] Compared with the amino acid sequence of wild-type rice EPSPS shown in SEQ ID NO:5, the rice EPSPS mutant I has a mutation of A155(138)T.

[0097] That is, relative to the wild-type rice EPSPS, the 155th amino acid residue of the rice EPSPS mutant I is mutated from G to A, which corresponds to the 138th position of E. coli EPSPS.

[0098] This embodiment also provides a rice EPSPS mutant I coding gene encoding the above rice EPSPS mutant I, the nucleotide sequence of which is shown in SEQ ID NO:15.

[0099] Both the rice EPSPS mutant I coding gene and the rice EPSPS mutant I provided in the examples of the present invention can be obtained by chemical synthesis.

Embodiment 2

[0101] This embodiment provides a plant EPSPS mutant, which is derived from rice and is rice EPSPS mutant II, which is obtained after mutation from wild-type rice EPSPS (amino acid sequence shown in SEQ ID NO: 5), and its amino acid sequence is as follows: Shown in SEQ ID NO:7.

[0102] Compared with the amino acid sequence of wild-type rice EPSPS shown in SEQ ID NO:5, the rice EPSPS mutant II has two mutations, G111(96)A and A155(138)T.

[0103] That is, if Figure 5 as shown ( Figure 5 Partial results of the amino acid sequence alignment of Escherichia coli EPSPS, rice EPSPS mutant II and wild-type rice EPSPS are shown, in the figure: Ec-EPSPS WT represents Escherichia coli EPSPS; Os-EPSPS M represents rice EPSPS mutant II; Os-EPSPS WT represents wild-type rice EPSPS), relative to wild-type rice EPSPS, the 111th amino acid residue of rice EPSPS mutant II is mutated from G to A, and this site corresponds to Escherichia coli EPSPS (SEQ ID NO :1) at the 96th position; the 1...

Embodiment 3

[0107] This embodiment provides a plant EPSPS mutant, which is derived from corn and is a corn EPSPS mutant, which is obtained by mutation of wild-type corn EPSPS (amino acid sequence shown in SEQ ID NO: 12), and its amino acid sequence is shown in SEQ ID NO: 12. ID NO:13.

[0108] With respect to the amino acid sequence of the wild-type corn EPSPS shown in SEQ ID NO: 12, the corn EPSPS mutant has K94(85)I, G105(96)A, P110(101)S and A149(138)T4 a mutation.

[0109] That is, if Image 6 as shown ( Image 6 Partial results of the amino acid sequence alignment of Escherichia coli EPSPS, maize EPSPS mutant and wild-type maize EPSPS are shown, in the figure: Ec-EPSPS WT represents Escherichia coli EPSPS; Z-EPSPSM represents maize EPSPS mutant; Z-EPSPS WT represents wild-type corn EPSPS), relative to the wild-type corn EPSPS, the 94th amino acid residue of the corn EPSPS mutant is mutated from K to I, which corresponds to the 85th position of Escherichia coli EPSPS; the 105th ami...

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Abstract

The invention discloses a plant EPSPS mutant with A138T mutation, and a coding gene and application of the mutant. Compared with escherichia coli EPSPS, the site, corresponding to the No. 138 site ofthe escherichia coli EPSPS, of the amino acid sequence of the plant EPSPS mutant provided by the invention is mutated as follows: A is larger than T. Through the mutation, the resistance of differentmutants of EPSPS of various plants to glyphosate is obviously improved, and meanwhile the catalytic activity of biological enzymes of the mutants is kept; the plants or recombinant strains, which aretransformed with the plant EPSPS mutants provided by the invention, can grow normally in the presence of the glyphosate; and the plant EPSPS mutant can be applied to not only culture of transgenic crops, but also culture of glyphosate-resistant non-transgenic plants such as rice, tobacco, soybeans, corn, wheat, cotton and sorghum, thereby having wide application prospects.

Description

[0001] This application claims the priority of the Chinese patent application with the application number 201711062151.X and the title "A Plant EPSPS Mutant Containing A138T Mutation and Its Encoding Gene and Application" submitted to the China Patent Office on November 2, 2017. The entire contents of which are incorporated by reference in this application. technical field [0002] The invention relates to the technical field of genetic engineering, in particular to a plant EPSPS mutant containing A138T mutation, its coding gene and application. Background technique [0003] Glyphosate is one of the most widely used herbicides in the world and has been used for nearly forty years. Glyphosate inhibits the activity of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). EPSPS catalyzes the synthesis of EPSP from PEP and shikimate-3-phosphate in the shikimate pathway, and finally synthesizes the aromatic amino acids tryptophan, phenylalanine and tyrosine. Glyphosate hinders t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54A01H5/00A01H6/46A01H6/54A01H6/82A01H6/60
CPCC12N9/1092C12N15/8275C12Y205/01019
Inventor 陈容邓龙群候青江卢远根欧倩冯小容李玲黄鑫胥南飞
Owner SICHUAN GEVOTO BIOTECH CO LTD
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