关键词: 细胞色素P450, 中华按蚊, CYP6P5, 鉴定, 生物信息学分析

Abstract:

Objective To identify CYP6P5 gene of Anopheles sinensis and to analyze its bioinformatic characteristics for further research on its role in insecticide resistance. Methods We searched the transcriptome dataset of An. sinensis for CYP6P5 cDNA sequence with bidirectional Blast, using the sequence of An. funestus CYP6P5 gene as query. The primary structure, hydrophobic region, transmembrane region, structural domain, and 3D structure of translated protein of the resultant sequence were analyzed using bioinformatic software. The phylogenetic relationship of CYP6P5 sequences of An. sinensis and representative mosquito species was constructed using maximum likelihood methods. Results A complete cDNA sequence of An. sinensis CYP6P5 was obtained by BLAST searching. The whole sequence was 1583 bp long with an open reading frame of 1527 bp, encoding 508 amino acids. The deduced amino acid sequence was found with characteristic conserved regions of superfamily P450 family 6 (CYP6). The gene was designated as CYP6P5 (GenBank No. KF358704). Bioinformatic prediction indicated that the molecular weight of CYP6P5 protein was 58.32×103, and the isoelectric point was 7.17. The sequence contained a predicted hydrophobic region spanning from locus 5 to 21, a transmembrane region from 5 to 22, a P450 domain from 36 to 505, and multiple enzyme active sites. Phylogenetic and similarity analyses of amino acid sequences showed that An. sinensis CYP6P5 had the closest phylogenetic relationship with CYP6P5s of An. funestus and An. gambiae, with similarity values of 89.4% and 89.0%, respectively. They formed a monophyletic group. Conclusion This study laid a foundation for further study on the molecular mechanism of An. sinensis CPY6P5s role in pyrethroid resistance.

Key words: Cytochrome P450, Anopheles sinensis, CYP6P5, Identification, Bioinformatic analysis