淡色库蚊嗅觉受体基因orco的克隆及组织表达谱分析

发布时间：2018-03-09 19:43

本文选题：淡色库蚊　切入点：嗅觉受体　出处：《中国病原生物学杂志》2017年09期 　论文类型：期刊论文

【摘要】：目的 Orco是嗅觉功能的重要组成元件。本研究旨在克隆淡色库蚊的orco基因,分析其编码蛋白的氨基酸序列和orco基因组织表达谱。方法利用RT-PCR技术克隆Cpporco基因,对其编码蛋白的氨基酸序列进行生物信息学分析;利用软件MEGA 5.0对获得的Cpporco进行系统进化树分析;利用HMMTOP version 2.0和TMHMM server version 2.0软件预测CppOrco的跨膜结构域;利用RT-PCR检测雌、雄淡色库蚊的触角、下颚须、喙、头、体中的orco基因表达水平。结果从淡色库蚊中克隆出一个嗅觉受体共表达基因Cpporco,编码区全长1 443bp,编码481个氨基酸,Cpporco与其他蚊种的orco的氨基酸一致性为83.63%,与同属于库蚊属的致倦库蚊Cqorco基因编码的氨基酸序列一致性为98.96%。系统进化树分析CppOrco与CqOrco的亲缘关系较近。CppOrco属于TM7的GPCR,其N末端在胞内,C末端在胞外。Cpporco基因在雌、雄蚊的不同组织中表达量不同,其中在雌蚊触角和喙高表达。结论成功克隆了淡色库蚊orco基因序列全长,其编码蛋白主要在雌、雄蚊触角和喙高表达,为进一步研究蚊虫的嗅觉系统及其功能奠定了基础。
[Abstract]:Orco is an important component of olfactory function. The purpose of this study is to clone the orco gene of Culex pipiens pallens, expression of orco gene and amino acid sequence analysis of its encoding protein. Methods using RT-PCR technique to clone Cpporco gene, the amino acid sequence of the protein encoding the biological information analysis; phylogenetic tree analysis using Cpporco software MEGA won 5 of 2 by HMMTOP version; and TMHMM server version 2 software to predict the transmembrane domain of CppOrco; RT-PCR is used to detect the female and male Culex tentacles, labialpalpi, beak, head, the expression level of orco gene in the body. Results from Culex pipiens pallens cloned from the olfactory receptor gene co expression Cpporco encoding region was 1 443bp, encoding 481 amino acids, amino acid consistency of Cpporco and other species of orco was 83.63%, and belong to the genus Culex Culex mosquito Cqorco Base Library Because of genetic relationship between amino acid sequence identity encoding for phylogenetic tree analysis of 98.96%. CppOrco and CqOrco is nearly.CppOrco belongs to TM7 GPCR, the N terminal in the intracellular C terminus of.Cpporco in extracellular gene expression in different tissues in female and male mosquitoes in different quantity, which is highly expressed in female antennae and beak. Objective to clone the full-length orco gene sequence of Culex pipiens pallens, its encoding protein mainly in female, high expression of the male mosquito tentacles and beak, which laid the foundation for the further study of mosquito olfactory system and its function.

【作者单位】：山东省医学科学院山东省寄生虫病防治研究所;
【基金】：国家自然科学基金项目(No.81471985,81672059) 山东省自然基金项目(No.ZR2014YL031,ZR2015YL032,ZR2015YL023) 山东省医学科学院医药卫生科技创新工程项目
【分类号】：R384.1

【相似文献】