白纹伊蚊电压门控钠离子通道基因克隆及其生物信息学分析

Cloning and bioinformatics analysis of voltage-gated sodium channel gene of Aedes albopictus

目的电压门控钠离子通道是拟除虫菊酯类杀虫剂作用的特异性靶标位点,获得白纹伊蚊电压门控钠离子通道编码基因cDNA序列,通过生物信息学技术分析其分子特征,为研究白纹伊蚊对拟除虫菊酯类杀虫剂产生靶标抗性的分子机制奠定基础。方法通过反转录-聚合酶链式反应(RT-PCR)技术克隆获得白纹伊蚊钠离子通道基因序列,利用生物信息学技术分析其生物学特征。结果白纹伊蚊钠离子通道基因全长为6 357 bp,编码2 118个氨基酸,等电点为5.06,蛋白相对分子质量为237×10^(3)。发现3个选择性剪接(m、h、d)和6个RNA编辑位点。BLAST序列分析表明,白纹伊蚊与埃及伊蚊钠离子通道基因具有很高的同源性,相似度高达96.30%,克隆所得白纹伊蚊钠离子通道基因序列符合昆虫钠离子通道基因的基本特征。结论克隆获得了白纹伊蚊钠离子通道编码基因全长cDNA序列,并分析了其生物信息学特征,有助于阐明抗性机制和新靶点的研究开发,特别是对拟除虫菊酯类杀虫剂产生的靶标抗性分子检测具有重要意义。

Objective Voltage-gated sodium channel(VGSC) is a specific target-site for pyrethroid insecticides. To obtain the cDNA sequence of the VGSC gene of Aedes albopictus and analyze its molecular characteristics using the bioinformatics techniques, and to provide a basis for investigating the molecular mechanism of Ae. albopictus developing target-site resistance to pyrethroid insecticides. Methods We cloned the VGSC gene sequence of Ae. albopictus by reverse transcription-polymerase chain reaction, and used bioinformatics techniques to analyze its biological features.Results The total length of the VGSC gene of Ae. albopictus was 6 357 bp, and encoded 2 118 amino acids, with an isoelectric point of 5.06 and a molecular weight of the protein of 237×10^(3). Three alternative splicing sites(m, h, and d)and six RNA editing sites were found. The basic local alignment search tool found that the VGSC genes of Ae. albopictus and Ae. aegypti were highly homologous, with the similarity of 96.30%, and the cloned VGSC gene sequence of Ae.albopictus had the typical characteristics of the sodium channel of insects. Conclusion The full-length cDNA sequence of the VGSC gene of Ae. albopictus is cloned and its bioinformatics characteristics are analyzed, which is helpful in clarifying resistance mechanism and developing new targets, especially for the molecular detection of target-site resistance to pyrethroid insecticides.