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为了初步探讨白纹伊蚊细胞色素P450 CYP6家族新基因分子进化机理,根据已获得的白纹伊蚊溴氰菊酯抗性株一新的细胞色素P450CYP6基因cDNA片段,设计反向引物,进行5’-cDNA末端快速扩增（rapid amplification of cDNA ends,RACE）;以正向引物进行3’-RACE。然后分别进行克隆、测序和鉴定。根据获得白纹伊蚊CYP6家族新成员全长cDNA序列,运用PC/GENE软件分析其5’-UTP区DNA序列相似性及编码区氨基酸残基相似性、构建系谱树。结果获得的3个全长序列中5’-UTP区DNA序列完全相同;编码区氨基酸残基相似性为3/497或4/497个氨基酸差异,而且此种差异均位于蛋白质功能非保守的基因区域;构建的系谱树显示CYP6N3基因与冈比亚按蚊CYP6N1-2亲缘关系最近,而与致倦库蚊的CYP6E1、CYP6F1亲缘关系较远。说明白纹伊蚊CYP6N3基因较致倦库蚊的CYP6E1、CYP6F1基因更为古老;白纹伊蚊CYP6N3各等位基因变异体是通过整个CYP6N3基因座的新近复制而产生。
In order to investigate the molecular evolution mechanism of CYP6 family of CYP6 gene from Aedes albopictus, a reverse transcription primer was designed based on a new cytochrome P450CYP6 cDNA fragment of Aedes albopictus resistant deltamethrin. ’-cDNA rapid amplification of cDNA ends (RACE); 3’-RACE with forward primer. Then were cloned, sequenced and identified. Based on the full-length cDNA sequence of the new member of CYP6 family of Aedes albopictus, the similarity of the 5’-UTP DNA sequence and the similarity of the amino acid residues in the coding region was analyzed by PC / GENE software to construct a phylogenetic tree. Results The DNA sequence of the 5’-UTP region in the three full-length sequences was identical. The amino acid residues in the coding region were 3/497 or 4/497 amino acids in length. All these differences were in the non-conserved genes The constructed phylogenetic tree showed that the CYP6N3 gene was closest to CYP6N1-2 of Anopheles gambiae and distantly related to CYP6E1 and CYP6F1 of Culex quinquefasciatus. The CYP6N3 gene of Aedes albopictus is more ancient than that of CYP6E1 and CYP6F1 genes of Culex quinquefasciatus. The alleles of CYP6N3 of Aedes albopictus are generated by the recent replication of CYP6N3 locus.