淡色庫蚊嗅覺受體基因orco的克隆及組織表達譜分析
發(fā)布時間:2018-03-09 19:43
本文選題:淡色庫蚊 切入點:嗅覺受體 出處:《中國病原生物學(xué)雜志》2017年09期 論文類型:期刊論文
【摘要】:目的 Orco是嗅覺功能的重要組成元件。本研究旨在克隆淡色庫蚊的orco基因,分析其編碼蛋白的氨基酸序列和orco基因組織表達譜。方法利用RT-PCR技術(shù)克隆Cpporco基因,對其編碼蛋白的氨基酸序列進行生物信息學(xué)分析;利用軟件MEGA 5.0對獲得的Cpporco進行系統(tǒng)進化樹分析;利用HMMTOP version 2.0和TMHMM server version 2.0軟件預(yù)測CppOrco的跨膜結(jié)構(gòu)域;利用RT-PCR檢測雌、雄淡色庫蚊的觸角、下顎須、喙、頭、體中的orco基因表達水平。結(jié)果從淡色庫蚊中克隆出一個嗅覺受體共表達基因Cpporco,編碼區(qū)全長1 443bp,編碼481個氨基酸,Cpporco與其他蚊種的orco的氨基酸一致性為83.63%,與同屬于庫蚊屬的致倦庫蚊Cqorco基因編碼的氨基酸序列一致性為98.96%。系統(tǒng)進化樹分析CppOrco與CqOrco的親緣關(guān)系較近。CppOrco屬于TM7的GPCR,其N末端在胞內(nèi),C末端在胞外。Cpporco基因在雌、雄蚊的不同組織中表達量不同,其中在雌蚊觸角和喙高表達。結(jié)論成功克隆了淡色庫蚊orco基因序列全長,其編碼蛋白主要在雌、雄蚊觸角和喙高表達,為進一步研究蚊蟲的嗅覺系統(tǒng)及其功能奠定了基礎(chǔ)。
[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.
【作者單位】: 山東省醫(yī)學(xué)科學(xué)院山東省寄生蟲病防治研究所;
【基金】:國家自然科學(xué)基金項目(No.81471985,81672059) 山東省自然基金項目(No.ZR2014YL031,ZR2015YL032,ZR2015YL023) 山東省醫(yī)學(xué)科學(xué)院醫(yī)藥衛(wèi)生科技創(chuàng)新工程項目
【分類號】:R384.1
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