白纹伊蚊细胞色素P450 CYP6家族基因多样性的研究(英文)

根据已获得的白纹伊蚊CYP6家族某成员cDNA序列片段AEDR ,设计基因特异性引物 ,以白纹伊蚊总RNA为模板 ,进行cDNA末端快速扩增 ,扩增产物经T -A克隆、测序。结果显示 :通过 5’ RACE获得 1个非全长cDNA序列 (GZS331 ) ,其与CYP6N1、CYP6N2的同源性分别为 59 8%和 59 1 % ,与CYP6N3v1 -v3同源性最高 ,达 83 9% - 84 3% ;通过 3’ RACE获得 6个非全长cDNA序列 ,其中来自抗性株的GZG0 33序列与CYP6N3v1 -v3的同源性达 98 2 % - 99 1 % ,而其余 3’ RACE克隆与CYP6N3v1 -v3的同源性则达 84 3% - 85 6%。上述所有非全长cDNA序列均与哺乳动物CYP3A1以及夜蛾CYP9A1有较高的同源性 ,分别为 2 3% - 36 1 %和 2 7 6% - 34 1 %。用PC/GENE软件所绘制的系统树显示出与同源性分析相一致的结果。所得非全长cDNA序列上报国际P450命名委员会进行统一的命名 ,并对蚊虫中细胞色素P450基因多样性及其形成原因进行了分析

STUDY ON THE DIVERSITY OF CYTOCHROME P450 GENES FROM AEDES ALBOPICTUS

Several pairs of specific primers according to the obtained cDNA sequence fragment from deltamethrin resistant Aedes albopictus were designed to amplify new CYP6 genes from total RNA of Aedes albopictus by rapid amplification of cDNA ends (RACE) technique. The products of RACE were cloned and selected for sequencing. The deduced amino acid sequences were subjected to homologous analysis. The results indicated that the identities of clone GZS331 sequence from 5'\|RACE products and clone GZG033 sequence from 3'\|RACE products to CYP6N3v1-v3 are 83 9%-84 3% and 98 2%-99 1% respectively; while the identities of the others from 3'\|RACE products to CYP6N3v1-v3 are 84 3%-85 6%. All of these obtained cDNA sequences have a higher homology to CYP3A1 in mouse and CYP9A1 in moth. The dendrogram constructed by PC/GENE software showed similar results to homologous analysis. These obtained sequences were submitted and named by the P450 Nomenclature Committee. The diversity of cytochrome P450 genes in Culicidae species was discussed.