pCW ori vector

The oriental tobacco budworm, Helicoverpa assulta, is a specialized pest that can cause serious damage to important crops such as peppers and tobacco. Various man-made pesticides have been used to control infestations of this pest. In order to understand how this pest responds to pesticides, the key players involved in pesticide transformation need to be identified. This study identified the P450 genes of the CYP6B subfamily in oriental tobacco budworm and revealed its expression pattern. Furthermore, the activity of HassCYP6B6 against 12 pesticides was explored using recombinant enzymes produced in E. coli.

In this study, pCW-HassCYP6B6 and pACYC-HaCPR were co-transformed into E. coli BL21 (DE3). A negative control without HassCYP6B6 was also established using pCW ori+ empty vector instead of pCW-HassCYP6B6. The starter culture was prepared by inoculating 5 mL of Luria Broth (LB) (containing 100 μg mL− 1 of both ampicillin and chloromycetin) with a single colony carrying both pCW-HassCYP6B6 (or pCWori+ empty plasmid as control) and pACYC-HaCPR plasmids and allowed growth overnight.

Figure 1. Contents of recombinant HassCYP6B6, NADPH-cytochrome P450 reductase (HaCPR) and cytochrome b5 (Hab5) in membrane preparations from E. coli cells. (Xiong, Tengfei, et al., 2024)