青南高原多房棘球蚴SBACT5基因的克隆和生物信息学分析  

Cloning and bioinformatics analysis of SBACT5 gene in Echinococcus multilocularis from Southern Qinghai Plateau

作　　者：何顺伟 李晓燕[3] 李洪清[4] 赵瑞雪[3] 魏晓星[5] 

机构地区：[1]青海大学生态环境工程学院,西宁810016 [2]青海大学附属医院教学管理部 [3]青海大学附属医院肝胆胰外科,西宁810016 [4]青海大学附属医院烧伤科,西宁810016 [5]青海大学医学院生化教研室,西宁810016

出　　处：《医学研究生学报》2017年第8期818-823,共6页Journal of Medical Postgraduates

基　　金：青海省科技厅项目(2016-ZJ-746)

摘　　要：目的胆汁酸钠协同转运蛋白在棘球绦虫发育过程中起着重要作用。文中研究克隆多房棘球蚴(Em)的胆汁酸钠协同转运蛋白5(Em SBACT5)基因,并对其编码蛋白进行生物信息学分析。方法采用RT-PCR技术扩增Em SBACT5基因并测序,采用生物信息学软件对编码蛋白的理化性质、亲/疏水性、跨膜域、翻译后修饰位点、结构域、二级结构、三级结构、亚细胞定位和生物学功能进行预测分析。结果扩增出654 bp的完整开放阅读框,其编码217个氨基酸,与公布的细粒棘球蚴(Eg)的Eg SBACT5基因核苷酸和氨基酸同源性分别达98%和96%。蛋白分析结果显示,Em SBACT5蛋白分子式为C1141H1797N273O284S11,相对分子量为24 240,p I为8.99。含有9个翻译后修饰位点和4处典型的结构域。α-螺旋、β-折叠、β-转角和无规则卷曲所占的比例分别为29.95%、31.80%、7.83%和30.41%。该蛋白为疏水性跨膜蛋白,主要定位于细胞质膜中,可能在物质转运和信号传递过程中发挥作用。结论成功克隆出Em SBACT5基因并获得其编码蛋白的的信息学特征,为包虫病防治提供基础信息。Objective Sodium-bile acid cotransporter plays an important role in the development of Echinococcus. The present study aimed to clone sodium bile acid cotransporter gene in Echinococcus multilocularis（Em SBACT5） and to analyze the bioinformatics of its coding protein. Methods Em SBACT5 gene was amplified by reverse transcription RCR（RT-PCR） technology and its nucleotide sequence was sequenced. Bioinformatics softwares were used to predict and analyze the physical and chemical properties,hydrophilicity/hydrophobicity,transmembrane domain,post-translational modification sites, structural domain, secondary structure, tertiary structure,subcellular localization and biological functions of the coding protein. Results The complete open reading frame was amplified with 654 bp in length,encoding 217 amino acids. The homology of the nucleic acid sequence and amino acid sequence of Em SBACT5 gene were 98% and 96% with the published SBACT5 in Echinococcus granulosus（Eg SBACT5） respectively. Protein analysis results showed that the molecular formula of Em SBACT5 protein was C1141H1797N273O284S11.Its relative molecular mass was 24 240 and isoelectric point was 8.99. There were 9 post-translational modification sites and4 typical domains. Alpha helical,β-sheet,β-turn and random coil accounted for 29.95%,31.80%,7.83% and 30.41%,respectively.This protein was a hydrophobic membrane protein and was mainly located in the cytoplasmic membrane,and it might play a role in the processes of material transport and signal transmission. Conclusion The Em SBACT5 gene was cloned successfully and the informatics characteristics of its coding protein were obtained,which provides basic information for prevention and control of echinococcosis.

关 键 词：多房棘球蚴 胆汁酸协同转运蛋白5 基因 克隆 生物信息学 分析 

分 类 号：R394[医药卫生—医学遗传学]