Bt protein in-vitro insect resistance identification method
A technology of bt protein and carrier, applied in the field of biological genetic engineering, to achieve the effects of not being moldy, maintaining integrity and biological activity, and expressing efficiently
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[0041] Embodiment 1 A method for identifying Bt protein against insects in vitro, comprising the following steps:
[0042] (1) Transform and synthesize the insect-resistant gene Bt ( figure 1) into the prokaryotic expression vector pET-28B (purchased from Shanghai Biological Engineering Co., Ltd.); the specific method is as follows:
[0043] Step 1: Digest pUC with restriction enzymes bt , the gel recovery kit recovers and purifies the Bt fragment;
[0044] Step 2: Digest pET28b with the same restriction enzyme, and recover and purify the digested pET28b fragment with the gel extraction kit;
[0045] Step 3: Ligate the two purified fragments, and construct the resulting prokaryotic expression plasmid named pET28b bt , identified by restriction endonuclease digestion, indicating that the vector was constructed correctly ( figure 2 );
[0046] (2) Transform the prokaryotic expression vector PET-28B containing the Bt gene into Escherichia coli BL21 (purchased from Shangh...
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