Standard plasmid molecular used for detecting transgenic soybean, corn and cotton and construction of the standard plasmid molecular

Abstract

The invention discloses a necessary standard plasmid molecular and a construction method of the standard plasmid molecular in the field of transgenic crop detection. The construction method comprises the following steps: modifying an octo-gene plasmid molecular pTLE8 (Patent Application No.201010286884.3), adding a fusion fragment of 3'-transformation event specific sequences of transgenic corn Bt176 strain and Mon810 strain and corn internal reference gene Hmg specific sequence, and modifying into a deca-gene plasmid molecular pTLH10 (Lec1, 35S, NOS, PAT, Sad1, Cry1, Ab / c, Bt176, Mon810 and Hmg). The standard plasmid molecular constructed by the invention can completely replace a positive standard sample, and is suitable for screening transgenic soybean GTS40-3-2 strain, transgenic corn Bt176 and Mon810 strains and transgenic Bt cottons, and qualitative and quantitative PCR analysis and detection of gene specificity and strain specificity.

Description

technical field [0001] The invention relates to a plasmid molecule in the technical field of molecular biology, in particular to a standard plasmid molecule for detection of transgenic soybean, corn and cotton and a construction method thereof. Background technique [0002] Since the commercialization of transgenic plants, the global GM planting area has continued to expand. From 1996 to 2009, the global GM crop planting area has reached 690 million hectares, and the GM crop planting area has increased by 67 times in the past 10 years. The GM crops grown are mainly GM soybean, GM corn and GM cotton. The GM soybean planted the largest area with 58.6 million hectares, accounting for 57% of the global GM crop area, followed by GM corn (25.2 million hectares, or 25%) and GM cotton (13.4 million hectares, or 13%). With the widespread cultivation of genetically modified crops, their safety issues have attracted widespread attention from the global public. In many countries, nota...

AI Technical Summary

Problems solved by technology

Genomic DNA extracted from positive standards (CRMs) is usually used as a positive control, but due to the biosafety of genetically modified crops and the limitations of existing technologies, it is difficult to obtain positive standards for genetically modified crops, so it is urgent to develop a new type of positive control Materials to meet evolving GMO detection needs

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