Abstract: 【Aim】 To identify genes of the CYP6Y subfamily in Anopheles sinensis, analyze their structure and characteristics, and deduce their possible functions. 【Method】 The CYP6Ys cDNA transcripts were retrieved and identified by two ways of Blasts using An. gambiae CYP6Y1 as query sequence against the transcriptome data of An. sinensis. The structure, characteristics and possible functions of the CYP6Y genes identified were analyzed using bioinformatics methods. 【Results】 Two genes of the CYP6Y subfamily were identified from An. sinensis transcriptome sequencing data, and named as AsCYP6Y1 (GenBank accession number: KF709397) and AsCYP6Y2 (GenBank accession number: KF709398) , respectively. Sequence analysis showed that AsCYP6Y1 and AsCYP6Y2 are 1 713 bp and 1 815 bp in length, encoding 502 and 526 amino acids, respectively. Gene structure analysis showed that the genes of this subfamily only contain one phase “1” intron and form a conserved synteny with other P450 genes. Protein structure prediction showed that the encoded proteins of these two genes contain five P450 characteristic sequences and six specific substrate binding sites, but have no signal peptide sequence, and are localized in cytoplasm. The 3D structural prediction showed that AsCYP6Y1 has 18 α-helix and 13 anti-parallel β-strands, while AsCYP6Y2 has 19 αhelix and 11 antiparallel βstrands. Two CYP6Y genes were also identified in An. darlingi using the same method. Phylogenetic analysis showed that AsCYP6Y1 and AsCYP6Y2 are grouped with CYP6Y1 and CYP6Y2 of other Anopheles species, respectively, with the bootstrap values greater than 90%. The Ka/Ks ratio analysis showed that the Ka/Ks ratio of AsCYP6Y1 and AsCYP6Y2 compared with CYP6Y1 and CYP6Y2 of other Anopheles species were all less than 1. The relative evolutionary rate test showed that the rates of subfamilies CYP6Y and CYP6M were significantly higher than those of the subfamily CYP6P in An. sinensis, while no significant difference between subfamilies CYP6Y and CYP6M was observed. 【Conclusion】 Two genes of the CYP6Y subfamily have been identified and characterized in An. sinensis and An. darlingi, which had earlier been found in two other Anopheles species (An. gambiae and An. funestus), suggesting the wide and specific existence of the genes of the CYP6Y subfamily in Anopheles.
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Key words: Anopheles sinensis;CYP6Y subfamily, gene sequence, bioinformatics analysis, gene structure, relative evolutionary rate