三种蚊虫COⅡ基因的克隆与序列分析

【作者】 黄朝晖；

【导师】 王金福；

【作者基本信息】 浙江大学 ， 遗传学， 2001， 硕士

【摘要】 蚊虫属双翅目(Diptera)长角亚目(Nematocera)蚊科(Culicidae)，是危害人畜的重要医学昆虫。中华按蚊(Anopheles sinensis)、白纹伊蚊(Aedes albopictus)、致倦库蚊(Culex quinquefasciatus)分别是按蚊属、伊蚊属和库蚊属的代表蚊种，均为最常见的蚊虫，在我国都有较广泛的分布。本文运用T-A克隆技术分别获得三种蚊虫的细胞色素C氧化酶亚基Ⅱ(COⅡ)基因重组质粒。测定和比较了中华按蚊、白纹伊蚊和致倦库蚊的COⅡ基因全序列，分析了三种蚊虫之间及与其它几种蚊虫的COⅡ基因序列间的差异及进化关系。并根据COⅡ基因序列构建了分子系统树，对这些蚊虫的进化年代及亲缘关系进行了讨论。结果显示：三种蚊虫COⅡ基因的核苷酸序列同源性为84.1％～87.9％，氨基酸序列的同源性为85.1％～89.5％；C+G含量为23.2％～24.9％，颠换频率高于转换频率。这三种蚊虫在COⅡ分子结构上，白纹伊蚊与致倦库蚊具有更近的亲缘关系；COⅡ基因序列用于蚊类种间系统发育的研究非常有效。更多还原

【Abstract】 Mosquitoes (Diptera: Culicidae), by nature of its association with problems of human and animal health, are medically important insects. Anopheles sinensis^ Aedes albopictus and Culex quinquefasciatus are all the most common mosquitoes in China. In this article, the cytochrome c oxidase subunit II gene (COII) in the mitochondrial DNA of the three mosquitoes was amplified by the polymerase chain reaction (PCR) technique, then was cloned into the pUCIS T-vector. The complete sequences of the three COII genes from Anopheles sinensis^ Aedes albopictus and Culex quinquefasciatus were sequenced and analyzed. The homology of the COII gene nucleotide and the deduced amino acid sequences of COII among three mosquitoes varied from 84.1% to 87.9% and from 85.1% to 89.5%, respectively. The C+G content of COII gene is about 23.2%~24.9%, and the frequency of transversion is higher than that of transition. The three mosquitoes were compared with others according to the COII gene, molecular phylogenetic tree was constructed base on the sequences of COII gene. The evolution history and relationships among those mosquitoes were discussed. Aedes albopictus and Culex quinquefasciatus are closely related species for their COII molecular level according to COII gene. The COII gene shows promise for resolving evolution and systematics of mosquitoes.更多还原