vip3a-1 gene of bacillus thuringiensis expression protein with high toxicity for asparagus caterpillar
A technology of Bacillus thuringiensis and beet armyworm, applied in the field of biochemistry, can solve problems such as poor control effect, threat to cotton production, and impact on cotton farmers' economic benefits
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[0025] 1. PCR amplification of genes:
[0026] The extracted Bt bacteria genomic DNA was amplified by PCR using the vip-5 / vip-3 primer pair, and the primer sequences are shown in Table 1. Referring to the N-terminal and C-terminal sequence homology of the coding region of vip3A gene published in GenBank, a pair of specific primers vip-5 / vip-3 were designed and synthesized, and NdeI and SalI restriction sites were introduced at the 5' end respectively (arrows indicated), for the amplification of the full-length gene.
[0027] Table 1: Primer sequences
[0028]
[0029] 2. PCR reaction system
[0030]
[0031]
[0032] 3. PCR reaction conditions
[0033] Table 2: PCR reaction conditions
[0034]
[0036] (1) Use a sterilized scalpel to cut off the gel containing the target DNA fragment and place it in a 1.5mL centrifuge;
[0037] (2) Add sol buffer A with 3 times the volume of sol, and place it in a water bath at 50°C for about 10 minut...
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