The invention discloses a standard
plasmid molecule necessary for transgenic
crop detection and a construction method thereof, and particularly discloses a standard
plasmid molecule for transgenic soybean detection and a construction method thereof. The standard
plasmid molecule comprises a soybean endogenous
gene Lectin-specific fragment, a 35S
promoter and NOS terminator-specific fragment, a transgenic soybean line A2704-12 foreign
gene PAT-specific fragment, and a transgenic soybean line GTS40-3-2 foreign
gene EPSPS-specific fragment. By analyzing the specific sequences of the above five genes, specific PCR (
polymerase chain reaction) primers are designed, and five
target gene fragments are obtained by amplification. By a fusion
PCR method, the five genes are fused to form a long fragment, and the long fragment is cloned to
plasmid vector pMDTM19-T Simple to obtain a recombinant plasmid molecule pTLE5. The standard plasmid molecule constructed in the invention can completely substitute for a positive
standard product, and is completely suitable for screening as well as structural-gene-specific qualitative and
quantitative PCR analysis and detection of transgenic soybean lines GTS40-3-2 and A2704-12. If a new transgenic material emerges, a new foreign gene fragment can be added to an original standard molecule, thus the detection requirements of increasing transgenic
crop materials are met.