Method for improving in-vitro expression of cytochrome P450 enzyme family

[0032] Example 1

[0033] 1. Construction and identification of recombinant transfer vector pFastBac1-CYP2A13 plasmid

[0034] pFastBac1-CYP2A13 is kept in this laboratory. After screening on ampicillin resistance plates, Escherichia coli containing recombinant plasmids was obtained, and bacterial liquid DNA was extracted to obtain recombinant transfer vector plasmid DNA. Use double enzyme digestion to verify pFastBac1-CYP2A13.

[0035] 2. Construction and identification of recombinant eukaryotic expression virus Ac-Bacmid-CYP2A13 DNA

[0036] The constructed pFastBac1-CYP 2A13 recombinant plasmid was transformed into competent cells of DH10Bac Escherichia coli. With the help of the helper plasmid of DH10Bac Escherichia coli, it was transposed by Tn7 transposon, and the gene fragments of each CYP2A13 were inserted into the shuttle vector Bacmid. The kanamycin, gentamicin and tetracycline tertiary antibodies were screened and blue-white spot screened to obtain recombinant Ac-Bacmid-CYP

[0042] Example 2

[0043] Compared with Example 1, it is the same except that the cofactor in step 4 is 0.05-0.6mM 5-ALA.

[0044] Example 3

[0045] Compared with Example 1, except that the cofactor in step 4 is 0.01-0.4mM Fe 3+ Except for the difference, everything else is the same.