淡色库蚊溴氰菊酯抗性相关基因ATP合酶g亚基cDNA克隆与序列分析

发布时间：2018-04-28 21:06

本文选题：ATP合酶g亚基 + 淡色库蚊　；参考：《中国病原生物学杂志》2017年08期

【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。
[Abstract]:Objective to clone the coding region of Culex pipiens pallens)ATP synthase g subunit gene from Culex pipiens pallens and analyze it by bioinformatics. To compare its expression in deltamethrin resistant and sensitive strains, the coding region of ATP synthase g subunit gene was amplified by PCR. The physicochemical properties, protein domain, transmembrane region, signal peptide and subcellular localization of ATP synthase g subunit gene encoding protein were predicted by bioinformatics analysis. Amino acid sequences of ATP synthase g subunits of Culex pipiens pallens and other species were compared and phylogenetic tree was constructed. The expression of ATP synthase g subunit gene in deltamethrin sensitive and resistant strains of Culex pipiens pallens was compared by real-time quantitative PCR. Results the coding region of ATP synthase g subunit gene of Culex pipiens pallens with long 306bp was cloned. The gene encodes 101 amino acids in GenBank accession number: KY783435. Bioinformatics analysis showed that the theoretical molecular weight unit of ATP synthase g gene encoding protein was 11 脳 10 ~ (-3) and the isoelectric point was 9.56. The 6-98 amino acid had ATP synthase g subunit domain, and no signal peptide or transmembrane region was found. Mitochondrial expression was the highest in subcellular localization prediction (39.1%). The expression of ATP synthase g subunit gene in deltamethrin resistant lines of Culex pipiens pallens was 3.14 times higher than that in sensitive strains by real-time quantitative PCR. Conclusion the coding region of ATP synthase g subunit gene of Culex pipiens pallens was obtained and analyzed by bioinformatics. It was confirmed that the gene was overexpressed in deltamethrin resistant lines.
【作者单位】：保定市第二中心医院血液肾病科;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;
【基金】：河北省高等学校科学技术研究项目(No.QN2016005)
【分类号】：R384.1

【相似文献】