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白纹伊蚊细胞色素P450 CYP6N3基因分子进化机制初探
A preliminary study on the molecular evolutionary mechanism of cytochrome P450 CYP6N3 gene from Aedes albopictus
【摘要】 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。
【Abstract】 To study the molecular evolutionary mechanism of cytochrome P450 CYP6N3 genes from Aedes albopictus. Method; A pair of specific primers according to an obtained cDNA fragment of cytochrome P450 CYP6 gene from deltamethrin-resistant Ae. albopictus were designed and used to amplify complete 5’ -end and 3’ -end by rapid amplification of cDNA ends(RACE) technique. The RACE products were cloned, sequenced and identified. The similarities of 5’ -UTR and amino acid residues in encoding region and dendrogram were analyzed and constructed by PC/GENE software, respectively. Results: The nucleotide sequences in 5’-UTR of the obtained full-length cDNAs are identical. There are 3 or 4 different amino acid residues out of 497 in their encoding regions, and these different residues are located in non-consevative region of P450 proteins. The constructed dendrogram shows that the CYP6N3 genes from Ae. albopictus have a closer relation to CYP6N1 and CYP6N2 genes from Anopheles gambiae than to CYP6E1 and CYP6F1 genes from Culex quinguefasciatus . Conclusions: The CYP6N3 alleles from Ae. albopictus were produced by recent replication of the whole CYP6N3 locus, and may be more ancient than CYP6E1 and CYP6F1 genes from Cx. quinquefasciatus .
【Key words】 Aedes albopictus cytochrome P450; CYP6N3; molecular evolution;
- 【文献出处】 广东寄生虫学会年报 , 编辑部邮箱 ,2000年00期
- 【分类号】R384.1
- 【被引频次】5
- 【下载频次】117