摘要
目的克隆淡色库蚊(Culex pipiens pallens)ATP合酶g亚基基因编码区序列并进行生物信息学分析,比较其在溴氰菊酯抗性品系和敏感品系中的表达方法利用PCR方法扩增ATP合酶g亚基基因编码区序列。利用生物信息学分析工具,预测ATP合酶g亚基基因编码蛋白的理化性质、蛋白结构域、跨膜区、信号肽和亚细胞定位;对淡色库蚊和其他物种的ATP合酶g亚基进行氨基酸序列多重比对,并构建系统发育树。通过实时定量PCR检测比较ATP合酶g亚基基因在淡色库蚊溴氰菊酯敏感品系和抗性品系中的表达。结果克隆出长306bp的淡色库蚊ATP合酶g亚基基因编码区序列(GenBank登录号:KY783435),该基因编码101个氨基酸。生物信息学分析ATP合酶g亚基基因编码蛋白理论分子质量单位(Mr)为11×10~3,等电点为9.56,在第6-98位氨基酸具有ATP合酶g亚基结构域,未发现信号肽和跨膜区,亚细胞定位预测以线粒体表达量最多(占39.1%)。实时定量PCR检测ATP合酶g亚基基因在淡色库蚊溴氰菊酯抗性品系中的表达量是敏感品系中表达量的3.14倍。结论获得淡色库蚊ATP合酶g亚基基因编码区序列,并进行了生物信息学分析,证实该基因在溴氰菊酯抗性品系中高表达。
Objectives To clone and bioinformatically analyze the coding sequence of the ATP synthase g subunit gene of Culex pipiens pallens and to compare its expression between the deltamethrin-susceptible and deltamethrin-resistant strains. Methods The coding sequence of the ATP synthase g subunit gene of Culex pipiens pallens was amplified with PCR.Tools for bioinformatic analysis were used to predict the physico-chemical properties,functional characteristics,transmembrane domains,signal peptides,and subcellular localization of the encoded protein,to align multiple amino acid sequences of the ATP synthase g subunit gene of Culex pipiens pallens and other species,and to construct a phylogenetic tree.Expression of the ATP synthase g subunit gene was compared between deltamethrin-susceptible and deltamethrin-resistant strains using quantitative real-time PCR. Results The coding sequence of the ATP synthase g subunit gene was cloned（GenBank accession no.：KY783435）.The sequence contained 306 nucleotides and encoded 101 amino acids with a theoretical relative molecular weight of 11 ku and an isoelectric point of 9.56 according to bioinformatic analysis.The ATP synthase g subunit was predicted to contain a structural domain（amino acids 6-98）of ATP-synt_G,to lack signal peptides and transmembrane domains,and to be expressed mostly in mitochondria（39.1%）.The level of expression of the ATP synthase g subunit gene in deltamethrin-resistant strains was 3.14 times that in deltamethrin-susceptible strains according to quantitative real-time PCR. Conclusion This study obtained the coding sequence of the ATP synthase g subunit gene of Culex pipiens pallens.This sequence was bioinformatically analyzed and its overexpression was verified in deltamethrin-resistant strains.
出处
《中国病原生物学杂志》
CSCD
北大核心
2017年第8期757-761,共5页
Journal of Pathogen Biology
基金
河北省高等学校科学技术研究项目(No.QN2016005)
关键词
ATP合酶g亚基
淡色库蚊
抗药性
溴氰菊酯
生物信息学
ATP synthase g subunit
Culex pipiens pallens
insecticide resistance
deltamethrin
bioinformatics
