淡色库蚊ATP合酶B亚基基因克隆和序列分析及其与溴氰菊酯抗性的关系

发布时间：2018-01-20 02:00

本文关键词： 淡色库蚊 ATP合酶B亚基抗药性溴氰菊酯生物信息学　出处：《昆虫学报》2017年08期 　论文类型：期刊论文

【摘要】：【目的】克隆淡色库蚊Culex pipiens pallens ATP合酶B亚基基因编码区序列,并进行生物信息学分析,研究其与溴氰菊酯抗性关系。【方法】通过PCR方法扩增ATP合酶B亚基基因编码区序列;利用生物信息学网站在线分析工具,预测ATP合酶B亚基基因编码蛋白的理化性质和功能特征;通过实时定量PCR方法比较ATP合酶B亚基基因在室内筛选的淡色库蚊溴氰菊酯敏感品系和抗性品系中的表达,并在现场种群溴氰菊酯敏感和抗性个体中做进一步验证。【结果】成功克隆淡色库蚊ATP合酶B亚基基因编码区序列(Gen Bank登录号:KY783434),长717 bp,编码238个氨基酸。生物信息学分析表明,其编码蛋白理论相对分子量为26.96 k D,等电点为8.97,在第70-231位氨基酸具有ATP合酶B亚基结构域,未发现信号肽和跨膜区。实时定量PCR结果显示,ATP合酶B亚基基因在室内筛选的淡色库蚊抗溴氰菊酯品系和现场种群溴氰菊酯抗性个体中表达量均上调。【结论】本研究获得了淡色库蚊ATP合酶B亚基基因编码区序列,进行了生物信息学分析,并证实其在溴氰菊酯抗性个体中高表达,为进一步研究该基因在蚊抗药性中的作用奠定了基础。
[Abstract]:[objective] to clone the coding region of subunit B of Culex pipiens pallens ATP synthase from Culex pipiens pallens and analyze it by bioinformatics. [methods] the coding region of ATP synthase B subunit gene was amplified by PCR. The physical and chemical properties and functional characteristics of ATP synthase B subunit gene encoding protein were predicted by online analysis tools of bioinformatics website. The expression of ATP synthase B subunit gene in susceptible and resistant strains of Culex pipiens pallens was compared by real-time quantitative PCR. The results were further verified in the susceptible and resistant individuals of the field population. [results] the coding region of ATP synthase B subunit gene of Culex pipiens pallens was cloned successfully (. Gen Bank: KY783434. The length of 717 BP encodes 238 amino acids. Bioinformatics analysis shows that the theoretical molecular weight of the encoded protein is 26.96 KD and the isoelectric point is 8.97. The ATP synthase B subunit domain was found at the 70-231 amino acid site, and no signal peptide or transmembrane region was found. The results of real-time quantitative PCR showed that there was no signal peptide or transmembrane domain. The expression of ATP synthase B subunit gene was up-regulated in the deltamethrin resistant strains of Culex pipiens pallens and in the field population. [conclusion] the ATP synthase B subunit of Culex pipiens pallens was obtained in this study. Sequence of gene coding region. Bioinformatics analysis was carried out, and its high expression in deltamethrin resistant individuals was confirmed, which laid a foundation for further study on the role of the gene in mosquito resistance.
【作者单位】：河北医科大学病原生物学教研室;南京医科大学病原生物学系;保定市第二中心医院血液肾病科;蚌埠市第三人民医院检验科;
【基金】：河北省高等学校科学技术研究项目(QN2016005)
【分类号】：R384.1
【正文快照】： 蚊可以传播疟疾、淋巴丝虫病、登革热、寨卡病毒病等多种蚊媒病,严重危害人类健康(Reiner etal.,2013;Mayer et al.,2017)。长期以来,化学防制在控制蚊媒、预防蚊媒病中具有重要作用(Scott etal.,2015)。但大量文献报道,蚊对一种或多种杀虫剂已经产生了不同程度抗性(梁军等,20

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