A transgenic rapeseed sd-rapeseed that aggregates commonly used screening targets and its application
A transgenic and rapeseed technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of differences in DNA extraction efficiency of transgenic components, error in detection sensitivity judgment, difficulty in accurately reflecting sensitivity, etc., to reduce labor costs and economic costs, Cost and labor saving, easy application effect
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[0048] Below in conjunction with accompanying drawing and embodiment the present invention is described in detail:
[0049] 1. Obtain T0 transgenic rapeseed through genetic transformation
[0050] 1.1, T-DNA vector construction
[0051] The 11 elements are proposed to include P-35S promoter, P-FMV35S promoter, P-NOS promoter, Bar gene, NPTII gene, HPT gene, Pmi gene, T-NOS terminator, T-35S terminator, T- The g7 terminator and T-e9 terminator were cloned into the binary vector pBI121. The T-DNA region of the pBI121 vector contains a reading frame of the NPTII gene, which is regulated by the P-NOS promoter and the P-NOS terminator, so only the DNA sequences of the remaining 8 elements need to be cloned. Cloning primers were designed based on the nucleotide sequences of the eight screening elements. Using the designed primers, the extracted genomic DNA was used as a template for PCR amplification. The PCR amplification system (50μl) contains: 1×KOD Plus buffer, MgSO4 1mmol / L...
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