猪带绦虫成虫EF-1基因生物信息分析及原核表达  

Bioinformatics analysis of function and prokaryotic expression of elongation factor-1 in Taenia. solium

作　　者：刘玉江[1] 戴佳琳[1] 黄江[1] 王宇[1] 

机构地区：[1]贵阳医学院多媒体形态学法医学及寄生虫学教研室,贵州贵阳550004

出　　处：《中国公共卫生》2011年第1期83-84,共2页Chinese Journal of Public Health

基　　金：国家自然科学基金(30760227);贵州省科技攻关项目(2008-3060)

摘　　要：目的分析和预测猪带绦虫成虫延伸因子（elongation factor-1,EF-1）基因及其编码蛋白的结构域特性,并进行原核表达。方法利用在线生物信息学网站及工具进行序列分析、预测其编码的延伸因子的理化特性、抗原表位、翻译后的修饰、功能域、亚细胞定位、拓扑结构、二级结构、三维空间构象等,将其编码区序列克隆岛原核表达载体pET-28 a（＋）上,测序鉴定重组质粒。结果该基因全长1 657 bp,编码区为186~1 533 bp,编码448个氨基酸,为全长基因;无跨膜区,具有多个磷酸化位点,蛋白的理化性质稳定,理论分子量为49 011.5 Da;没有质体、线粒体定位序列;十二烷基-聚丙烯酰胺凝胶电泳（SDS-PAGE）结果表明,目的基因在大肠埃希菌BL21~DE3中表达成功。结论发现猪带绦虫成虫延伸因子基因,成功构建重组原核表达质粒。Objective To analyze and predict structural characteristics of elongation factor-1 in Teania.solium with bioinformatics and to detect its prokaryotic expression.Methods By online analysis with bioinformatics websites and software package,the physical-chemical characteristics,modification sites after translation,domains,subcelluar location,topological structure,second structure,and 3D structure of elongation factor-1（EF-1） were analyzed.The EF-1 gene was inserted into the prokaryotic expression vector pET-28a（＋） and then was amplified with PCR and sequenced.Results A novel cDNA sequence EF-1 with 1657 pb coding 488 amino acids and a molecular weight of 49011.5 was identified.PCR,double enzyme digestion and DNA sequencing indicated that pET-28a（＋） and EF-1 recombinant plasmid were successfully constructed.Sodium dodecyl sulfate polyaerylamide gel electrophoresis（SDS-PAGE） results showed that the gene was expressed in Escherichia coli BL21/DE3.Conclusion A novel gene of Taenia.solium is successfully identified and expressed with prokaryotic vector.

关 键 词：猪带绦虫 生物信息 原核表达 

分 类 号：R532.3[医药卫生—内科学] Q786[医药卫生—临床医学]