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A technology for transformants and herbicides, which is applied in biochemical equipment and methods, determination/inspection of microorganisms, etc., can solve the problem of no herbicide-resistant cotton-specific fragment sequences, etc., and achieves high practical value, experimental conditions, and sensitivity. high effect
Inactive Publication Date: 2012-11-14
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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Problems solved by technology
[0005] After searching the public literature and patents, it was found that there is no report on the transformant-specific fragment sequence of herbicide-resistant cotton LLCOTTON25
Method used
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Embodiment 1
[0023] Example 1 Cloning of specific sequences of transgenic herbicide-resistant cotton LLCOTTON25 transformants
[0026] The genetically modified herbicide-resistant cotton LLCOTTON25 (Bayer Crop Science Company) is preserved in this laboratory and can be distributed to the public for experimental research.
[0029] Other biochemical reagents are imported subpackages or domestic analytical pure. Primers were synthesized by Beijing Aoke Biotechnology Co., Ltd.
[0069] The transformant-specific PCR method of the present invention is applied to the identification of transgenic herbicide-resistant cotton LLCOTTON25
[0072] The genetically modified herbicide-resistant cotton LLCOTTON25 (Bayer Crop Science Company) is preserved in this laboratory and can be distributed to the public for experimental research.
[0073] 8 cotton materials, purchased from the market.
[0074] 1.2 Enzymes and Reagents
[0075] Molecular biology reagents, such as dNTPs, Taq DNA polymerase, Marker, etc. were purchased from Dalian Baobio Bioengineering Co., Ltd. Other biochemical reagents are imported subpackages or domestic analytical pure. Primers were synthesized by Beijing Aoke Biotechnology Co., Ltd.
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Abstract
The invention relates to a PCR (PolymeraseChain Reaction) method for identifying transistance phytocide cotton LLCOTTON 25 transformant, belonging to the field of transgenosis plant detection. In the invention, a transformant specificity sequence of the transgenosis phytocide cotton LLCOTTON 25 is obtained through a Genome walking method, wherein one is the specificity side sequence in the inserting area of an exogenous recombination carrier 5' end, and the other is a specificity side sequence in the inserting area of a xogenous recombination carrier 3'. A primer pair is designed according to any one of the two side sequences for detecting the transgenosis cotton breed LLCOTTON25 and the cotton breed using the cotton as the parent by PCR detection.
Description
technical field [0001] The invention belongs to the field of detection of transgenic plants, in particular to a PCR method for identifying LLCOTTON25 transformants of transgenic herbicide-resistant cotton Background technique [0002] my country is one of the major planting countries of genetically modified crops, and its planting area ranked sixth in the world in 2009. The transgenic crops commercially planted in my country are mainly transgenic insect-resistant cotton and transgenic papaya resistant to papaya ringspot virus. Among them, Bt geneinsect-resistant cotton is the transgenic crop with the largest planting area in my country. In 2009, the planting area reached 3.7 million hectares, accounting for nearly 70% of the total cotton planting area in my country (James, Clive. 2009. Global Status of Commercialized Biotech / GM Crops: 2009. ISAAA Brief No. 41. ISAAA: Ithaca, NY.). The transgenic cotton planted in my country is mainly insect-resistant transgenic cotton. In...
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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 谢家建孙爻张永军彭于发
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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