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Plant EPSPS mutant containing L195P and S247G as well as encoding gene and application thereof

A technology for coding genes and mutants, applied in plant EPSPS mutants and their coding genes and application fields, can solve the problems of low acceptance of transgenic crops, achieve the effects of maintaining the catalytic activity of biological enzymes, improving resistance, and broad application prospects

Active Publication Date: 2018-12-18
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. Several crops such as soybeans and cotton

Method used

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  • Plant EPSPS mutant containing L195P and S247G as well as encoding gene and application thereof
  • Plant EPSPS mutant containing L195P and S247G as well as encoding gene and application thereof
  • Plant EPSPS mutant containing L195P and S247G as well as encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0117] The soybean EPSPS mutant provided in this example is named G1, and its amino acid sequence is shown in SEQ ID NO.4.

[0118] The soybean EPSPS mutant G1 provided in this example is composed of wild-type soybean EPSPS (named as G0, the amino acid sequence is shown in SEQ ID NO.2) corresponding to the No. The 195th position (the mutation site is located at the 212th position of the wild-type soybean EPSPS) is obtained by mutating the amino acid residue L to P.

[0119] The amino acid sequence alignment of soybean EPSPS mutant G1, wild-type soybean EPSPS and Escherichia coli EPSPS is as follows: figure 1 As shown, in the figure: the position indicated by the arrow is the mutation site, and Ec-EPSPS WT represents Escherichia coli EPSPS.

[0120] In addition, this embodiment also provides a nucleic acid molecule (ie, coding gene) encoding soybean EPSPS mutant G1, the base sequence of which is shown in SEQ ID NO.3.

[0121] Both the soybean EPSPS mutant G1 and the nucleic a...

Embodiment 2

[0123] The soybean EPSPS mutant provided in this example is named G3, and its amino acid sequence is shown in SEQ ID NO.8.

[0124] The soybean EPSPS mutant G3 provided by the present embodiment is composed of the wild-type soybean EPSPS (named as G0, the amino acid sequence is shown in SEQ ID NO.2) corresponding to the No. The 195th position (the mutation site is located at the 212th position of wild-type soybean EPSPS) is mutated from amino acid residue L to P and the 101st position (this site corresponds to the 114th position of wild-type soybean EPSPS) is mutated from amino acid residue P to S get.

[0125] The amino acid sequence alignment of soybean EPSPS mutant G3, wild-type soybean EPSPS and Escherichia coli EPSPS is as follows: figure 2 As shown, in the figure: the position indicated by the arrow is the mutation site, and Ec-EPSPS WT represents Escherichia coli EPSPS.

[0126] In addition, this embodiment also provides a nucleic acid molecule encoding soybean EPSPS...

Embodiment 3

[0129] The soybean EPSPS mutant provided in this example is named G4, and its amino acid sequence is shown in SEQ ID NO.10.

[0130] The soybean EPSPS mutant G4 provided by the present embodiment is composed of the wild-type soybean EPSPS (named as G0, the amino acid sequence is shown in SEQ ID NO.2) corresponding to the E. coli EPSPS (the amino acid sequence is shown in SEQ ID NO.41). The 247th position (this position corresponds to the 265th position of the wild-type soybean EPSPS) is obtained by mutation of the amino acid residue S to G.

[0131] The amino acid sequence alignment results of soybean EPSPS mutant G4, wild-type soybean EPSPS and Escherichia coli EPSPS are as follows: image 3 As shown, in the figure: the position indicated by the arrow is the mutation site, and Ec-EPSPS WT represents Escherichia coli EPSPS.

[0132] In addition, this embodiment also provides a nucleic acid molecule encoding soybean EPSPS mutant G4, the base sequence of which is shown in SEQ I...

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Abstract

The invention discloses a plant EPSPS mutant containing L195P and S247G as well as encoding gene and application thereof and relates to the technical field of gene engineering. Compared with Escherichia coli EPSPS, an amino acid sequence of the plant EPSPS mutant disclosed by the invention has mutant L195P corresponding to 195th site of the Escherichia coli EPSPS and / or has mutant S247G corresponding to 247th site of the Escherichia coli EPSPS. Mutation of any one site of the two sites or simultaneous mutation of the two sites can give or improve resistance of the plant EPSPS mutant to glyphosate. Plants or recombinant bacteria for converting the plant EPSPS mutant can normally grow under the condition of glyphosate existence. The plant EPSPS mutant not only can be applied to culturing transgenic crops, but also can be applied to culturing glyphosate resisting non-transgenic plants like rice, tobacco, soybean, corn, wheat, cotton, sorghum and the like.

Description

[0001] This application claims the priority of a Chinese patent application with the application number 201711416358.2 and titled "A Plant EPSPS Mutant Containing L195P and S247G Mutations and Its Encoding Gene and Application" submitted to the China Patent Office on December 25, 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 L195P and S247G mutations, its encoding 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. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54C12N15/82A01H5/00A01H6/00
CPCC12N9/1085C12N15/8275C12Y205/01019C12N9/1092
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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