Plant EPSPS mutant containing A138X1 and L195X2 mutation and coding genes and application of plant EPSPS mutant

A technology of L195X2 and A38X1, applied in the field of genetic engineering, 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

Inactive Publication Date: 2019-04-02
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 A138X1 and L195X2 mutation and coding genes and application of plant EPSPS mutant
  • Plant EPSPS mutant containing A138X1 and L195X2 mutation and coding genes and application of plant EPSPS mutant
  • Plant EPSPS mutant containing A138X1 and L195X2 mutation and coding genes and application of plant EPSPS mutant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0113] The rice EPSPS mutants provided in this example are named as OsC, OsD, OsE, OsI, OsL, OsM, OsN and OsS, and their amino acid sequences are respectively as SEQ ID NO.49, SEQ ID NO.50, SEQ ID NO.51 , SEQ ID NO.52, SEQ ID NO.53, SEQ ID NO.54, SEQ ID NO.55, SEQ ID NO.56.

[0114] The rice EPSPS mutant provided in this example is composed of wild-type rice EPSPS (named OsEPSPS_WT, the amino acid sequence is shown in SEQ ID NO.48) corresponding to the 138th position of Escherichia coli EPSPS (the amino acid sequence is shown in SEQ ID NO.47). The position (the mutation site is located at the 155th position of the wild-type rice EPSPS) is obtained by mutating the amino acid residue A to C, D, E, I, L, M, N or S.

[0115] The amino acid sequence alignment of rice EPSPS mutants OsC, OsD, OsE, OsI, OsL, OsM, OsN and OsS, wild-type rice 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-...

Embodiment 2

[0119] The rice EPSPS mutants provided in this example are named OAC and OASC, and their amino acid sequences are shown in SEQ ID NO.58 and SEQ ID NO.60; the soybean EPSPS mutants provided in this example are named GAC and GASN, and their amino acid sequences are As shown in SEQ ID NO.63 and SEQ ID NO.65; the provided wheat EPSPS mutant is named TASC, and its amino acid sequence is shown in SEQ ID NO.68.

[0120] The rice EPSPS mutants OAC and OASC provided in this example are composed of wild-type rice EPSPS (OsEPSPS_WT, the amino acid sequence is shown in SEQ ID NO.48) corresponding to the Escherichia coli EPSPS (the amino acid sequence is shown in SEQ ID NO.47). The 138th position (the mutation site is located at the 155th position of wild-type rice EPSPS) is mutated from amino acid residue A to C and the 96th position (this site corresponds to the 111th position of wild-type rice EPSPS) is mutated from amino acid G to A And the 101st position (this position corresponds to ...

Embodiment 3

[0125] The rice EPSPS mutant provided by the present embodiment is named as RA, RC, RE, RF, RG, RH, RK, RM, RN, RQ, RR, RS, RT, RV and RY, and its amino acid sequence is respectively as SEQ ID NO .69, SEQ ID NO.70, SEQ ID NO.71, SEQ ID NO.72, SEQ ID NO.73, SEQ ID NO.74, SEQ ID NO.75, SEQ ID NO.76, SEQ ID NO.77, Shown in SEQ ID NO.79, SEQ ID NO.80, SEQ ID NO.81, SEQ ID NO.82, SEQ ID NO.83, SEQ ID NO.84.

[0126] The rice EPSPS mutant provided in this example is composed of wild-type rice EPSPS (named OsEPSPS_WT, the amino acid sequence is shown in SEQ ID NO.48) corresponding to the 195th position of Escherichia coli EPSPS (the amino acid sequence is shown in SEQ ID NO.47). position (the mutation site is located at the 214th position of wild-type rice EPSPS) is mutated from amino acid residue L to A, C, E, F, G, H, K, M, N, Q, S, T, V or Y get.

[0127] The amino acid sequence alignment of rice EPSPS mutants RA, RC, RE, RF, RG, RH, RK, RM, RN, RQ, RR, RS, RT, RV and RY, wild-t...

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Abstract

The invention discloses a plant EPSPS mutant containing A138X1 and L195X2 mutation and coding genes and application of the plant EPSPS mutant, and relates to the technical field of genetic engineering. Compared with escherichia coli EPSPS, the plant EPSPS mutant disclosed by the invention has the advantage that the amino acid sequence of the plant EPSPS mutant corresponds to the 138th site havingmutation A38X1, and/or the 195th site having mutation L195X2, of the escherichia coli, wherein X1=C, D, E, I, L M, N or S, and X2=A, C, E, F, G, H, K, M, N, Q, R, S, T, V or Y. When either one of twosites is mutated, or two sites are in mutation at the same time, the resistance of the plant EPSPS mutant to glyphosate can be given or improved. Strains or recombinant strains converting the plant EPSPS mutant can normally grow under the condition that the glyphosate exists.

Description

technical field [0001] The present invention relates to the technical field of genetic engineering, in particular, to an A138X-containing 1 and L195X 2 Mutated plant EPSPS mutants and their encoding genes and applications. Background technique [0002] 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 the synthesis of aromatic amino acids, which in turn affects the normal growth of plants and eventually leads to death. [0003] At present, the main method of cultivating glyphosate-resistant varieties is to introduce glyphosate-resistant genes from bacteria into crops by means of genetic engineering, so as to breed new varie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/10C12N15/54C12N15/82A01H5/00
CPCC12N9/1092C12N15/8275C12Y205/01019
Inventor 陈容候青江邓龙群张震胥南飞
Owner SICHUAN GEVOTO BIOTECH CO LTD
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