PSOY19-ZM2 vector as well as preparation method and application thereof
A technology of psoy19-zm2 and 1.psoy19-zm2, applied in the field of genetically modified soybeans, can solve the problems of reducing the content of herbicide-resistant protein in seeds, not being able to fully recognize the food safety of genetically modified soybeans, and high accumulation, so as to improve the ability of herbicide resistance , Alleviate safety concerns and increase production
Active Publication Date: 2021-08-03
ZHEJIANG UNIV
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Problems solved by technology
[0003] The previously obtained herbicide-resistant transgenic soybeans are all constitutively expressed, that is, the resulting transgenic soybean seeds also have EPSPS protein accumulation, and the accumulation is relatively high, which makes the public unable to fully recognize the food safety of genetically modified soybeans. Therefore, the inventors Developed the pSOY19-ZM2 vector, its preparation method and application, and reduced the herbicide-resistant protein content in seeds under the condition of ensuring high-efficiency herbicide resistance
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[0022] In this example, the pSOY19-ZM2 vector was first constructed. The pSOY19-ZM2 vector can increase soybean herbicide resistance and reduce the content of herbicide-resistant protein in soybean seeds. The pSOY19-ZM2 vector can be used to increase soybean herbicide resistance and reduce soybean In products with herbicide resistant protein content in seeds.
[0023] In this embodiment, the promoter sequence of 2171bp upstream of the initiation codon of the GmCRB1 gene is first subjected to PCR amplification, and the amplification primers are as follows:
[0024] PGmCRB1-F:
[0025] cccgggtaccgagctcGAATTCTCTTATAATAACCCTGTTAAC,
[0026] PGmCRB1-R:
[0027] gagactggtgatGAATTCGTTTCGTTCCACACAGTGC; the amplified PGmCRB1 fragment is obtained, and the nucleotide sequence of PGmCRB1 is shown in SEQ ID NO: 1, which is reserved; the 35S promoter in the plant expression vector pSOY19 vector is excised with EcoRI and NcoI double enzymes; then the PGmCRB1 fragment is excised with The 3...
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The invention relates to the technical field of transgenic soybeans, and discloses a pSOY19-ZM2 vector. The pSOY19-ZM2 vector is formed by connecting a green tissue specific promoter PGmCRB1 and a pSOY19 vector of which a 35S promoter is cut off; the invention also discloses a preparation method of the vector. The invention also discloses application of the vector. According to the vector, the expression of the herbicide-resistant gene G10-EPSPS is started by using a soybean green tissue specific promoter, so that herbicide-resistant protein is expressed in green tissues such as soybean leaves and the like, and is low-expressed or zero-expressed in seeds, so that the worry of the public on the safety of transgenic soybeans is relieved, and the herbicide-resistant gene G10-EPSPS is obtained. According to the application of the invention, a more direct and effective way is opened up for cultivating transgenic crops with efficient glyphosate resistance and reduced glyphosate-resistant protein in seeds, and the transgenic soybeans are improved in the acceptability of the public to the transgenic soybeans, the herbicide resistance of the soybeans is also improved, and the yield of the soybeans is improved. The application of the invention provides a more direct and effective way for cultivating the transgenic crops with efficient glyphosate resistance and reduced glyphosate-resistant protein in the seeds.
Description
technical field [0001] The invention relates to the technical field of transgenic soybeans, in particular to pSOY19-ZM2 vector, its preparation method and application. Background technique [0002] Soybean (GLycine max.Merr.) is an important oil crop and national protein source, which is of great significance in agricultural production. In the process of soybean growth, diseases, insect pests and weeds have seriously affected the yield and quality of soybeans. Compared with foreign soybean production, the competitiveness of soybean production in my country is very weak, and more than 85% depend on imports, which is a "stuck neck" problem. Therefore, there is an urgent need to increase the yield and quality of soybeans in my country and improve the international competitiveness of soybean production by cultivating high-quality resistant strains. [0003] The previously obtained herbicide-resistant transgenic soybeans are all constitutively expressed, that is, the resulting t...
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IPC IPC(8): C12N15/82C12N15/54C12N15/66A01H5/10A01H6/54
CPCC12N15/8223C12N15/8274C12N15/66C12N9/1092C12Y205/01019
Inventor 李林朱佳美王守冬徐恬恬寿惠霞
Owner ZHEJIANG UNIV