Studies on molecular mechanism of Anopheles anthropophagus to deltamethrin Ⅰ. Cloning and sequence analysis of CYP6 gene cDNA fragments from Anopheles anthropophagus / 中国血吸虫病防治杂志

ABSTRACT

Objective To obtain cDNA fragment of CYP6 gene from Anopheles anthropophagus. Methods The degenerate RT-PCR technique was used to amplify the CYP6 mRNA from susceptible and deltamethrin-resistant strains of An.anthropophagus. The specific cDNA fragments were sequenced and analyzed. Results The 250 bp specific cDNA fragments of CYP6 gene from both susceptible and resistant strains were cloned and sequenced by the Direct Cloning Method. Total of 10 sequences, including 2 from susceptible strain and 8 from resistant strain, were obtained. Compared with P450 sequence databases from GenBank, all of the 10 sequences obtained were new P450 sequence and were lodged in GenBank (Accession numbers AY273927-AY273936). According to being checked up by Nomenclature Committee of Cytochrome P450, 7 of them were confirmed as the new CYP6 genes and the others were alleles and all of them belonged to the subfamilies CYP6Z, CYP6P, CYP6N and CYP6M in CYP6 family, respectively. Conclusion The cloned 10 sequences are gene fragments from members of CYP6 family.[