棉铃虫细胞色素P450家族 CYP6AE14 基因克隆、序列分析及蛋白原核表达

采用同源克隆技术克隆石河子棉区棉铃虫抗性品系的P450家族解棉酚基因 CYP6AE14 。克隆获得的目的基因全长1 581 bp，编码526个氨基酸。序列分析表明，该抗性品系的 CYP6AE14 有1个氨基酸发生突变，即Y25F；其与烟草天蛾的CYP6AE32同源性较高，达60%。利用原核表达载体pET28 a在大肠杆菌中成功表达棉铃虫P450 CYP6AE14 基因，为进一步的基因功能验证奠定基础。

Cloning, Sequence Analysis and Prokaryotic Expression of Cytochrome P450 CYP6AE14 Gene from Helicoverpa armigera

In this study, a gossypol detoxification P450 gene CYP6AE14 from resistant strains of cotton bollworm in the Shihezi reclamation area, was isolated by the homology cloning techniques. The 1 581 bp full length orf of Helicoverpa armigera P450 CYP6AE14 encodes 526 amino acid residues. Sequence analysis shows that a single amino acid change in the sequence of resistant strains of cotton bollworm: Y25F. The cDNA deduced amino acid sequence of CYP6AE14 was similar to that of CYP6AE32 from Manduca sexta, which shows 60% amino acid sequence identity.The gene can be successfully expressed by the recombinant prokaryotic expression vector in E.coli, which laid the foundation for further gene function identification.