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Expression vector of glyphosate resistance genes GR79 and GAT and application

A technology of glyphosate resistance and expression vectors, which can be applied in the direction of vectors, applications, nucleic acid vectors, etc., and can solve problems such as the impact of herbicides on the growth and development of crops

Pending Publication Date: 2021-05-28
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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Problems solved by technology

[0004] However, due to the non-selective characteristics of herbicides, it will inevitably affect the growth and development of crops during use.

Method used

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  • Expression vector of glyphosate resistance genes GR79 and GAT and application
  • Expression vector of glyphosate resistance genes GR79 and GAT and application
  • Expression vector of glyphosate resistance genes GR79 and GAT and application

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Embodiment Construction

[0029] The present invention will be further described in detail below in conjunction with the examples.

[0030] The biological materials involved in the following are all preserved in the applicant's laboratory and can be distributed to the outside world.

[0031] 1. Codon modification and optimization of GR79 and GAT genes

[0032] The GR79 gene was isolated and cloned from the metagenome of soil bacteria heavily polluted by glyphosate, and has independent intellectual property rights (patent number: ZL 200710177090.1). The coding sequence of the GR79 gene has 1338bp, the nucleotide sequence is shown in SEQID NO.1, and the EPSPS enzyme encoded by it consists of 445 amino acids. The coding sequence of the GR79 gene is optimized according to the codons preferred by plants. The GC content of the original GR79 gene is 45.85%, and the GC content of the optimized GR79 gene is 64.56%. The optimized nucleotide sequence is shown in SEQ ID NO.3. The GAT gene is obtained by construc...

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Abstract

The invention relates to an expression vector of glyphosate resistance genes GR79 and GAT and application of the expression vector. The invention provides a novel DNA sequence synthesized by optimizing codons of a gene GR79and a gene GATwith high glyphosate tolerance and a plant expression vector pCGG constructed by using double genes, and research results prove that the gene GR79 and the gene GAT in different tissues of the transgenic corn transferred into the plant expression vector can be normally transcribed and translated, the expression quantity of the GR79 protein in the leaves in the seedling stage is 733.15-2203.99 ng / g, the expression quantity of the GAT protein is 101.56-345.84 ng / g, and the two proteins have high expression quantity in corn. Results show that the GR79 and GAT transgenic corn has high resistance to the target herbicide glyphosate, and the expression vector has important significance for synergistically expressing and cultivating the high-glyphosate-resistance plant by using the two genes to transform the plant and applying the high-glyphosate-resistance plant.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to an expression vector of two glyphosate resistance genes, GR79 and GAT, and their combined application in plant glyphosate tolerance. Background technique [0002] There are more than 250 kinds of main weeds in my country's farmland, with a distribution area of ​​more than 40 million hectares, of which more than 10 million hectares of farmland have been severely damaged. In our country, the average annual crop yield reduction due to weeds is about 13%, and the direct economic loss accounts for 10% to 20% of the total crop output value. The use of chemical methods to control weeds has become an indispensable part of modern agriculture. [0003] Glyphosate is a broad-spectrum biocidal and systemic herbicide. With its broad-spectrum, high efficiency, and environmental friendliness, it has become the most widely used pesticide in the world. The shikimate pathway is an important pathway f...

Claims

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Application Information

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IPC IPC(8): C12N15/84C12N15/54A01H5/00A01H6/46
CPCC12N9/1092C12N9/1029C12N15/8275C12Y205/01019C12N2800/22
Inventor 李圣彦郎志宏李香银林敏陆伟李鹏程张明俊
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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