Pyrethroid insecticide hydrolytic enzyme genes
A pyrethroid insecticide and hydrolase technology, applied in hydrolase, genetic engineering, plant genetic improvement and other directions, can solve problems such as unrealistic, and achieve the effect of improving product quality and increasing added value
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[0021] 1. Cloning of pyrethroid hydrolase gene
[0022] 1.1 Extraction of total DNA from Sphingomonas sp.JQ1-11 (GenBank registration number DQ132883)
[0023] The high-purity and high-molecular-weight JQ1-11 total DNA was extracted by CTAB method, dissolved in TE (pH8.0), and stored at -20°C. For specific methods, refer to "Refined Molecular Biology" edited by F. Osper et al. Experiment Guide".
[0024] 1.2PUC118 (BamHI) was purchased from Bao Biological Engineering (Dalian) Co., Ltd.
[0025] 1.3 Digestion of total DNA The total DNA of Sphingomonas sp. JQ1-11 was partially digested with Sau3AI.
[0026] 1.4 Recovery of DNA
[0027] The total DNA after enzyme digestion was purified by electrophoresis (TAE buffer) and recovered using axygen biosciences (China) recovery kit. The recovered DNA was dissolved in 10 mmol / L Tris Cl (PH8.0) and placed in- Store at 20°C.
[0028] 1.5 Enzyme linkage to establish the following reaction system:
[0029] PUC118 (BamHI) 0.1 μg
[00...
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