阴道毛滴虫TvRab11C基因的克隆及原核表达  

Cloning and prokaryotic expression of TvRab11C gene of Trichomonas vaginalis

作　　者：刘畅[1] 丁鹤[1] 宫鹏涛[1] 李建华[1] 李淑红[1] 李赫[1] 张国才[1] 张西臣[1] 

机构地区：[1]吉林大学农学部畜牧兽医学院,长春130062

出　　处：《中国生物制品学杂志》2012年第7期834-836,共3页Chinese Journal of Biologicals

基　　金：艾滋病和病毒性肝炎等重大传染病防治(2008ZX10401-B)

摘　　要：目的克隆并原核表达阴道毛滴虫TvRab11C(G3 Ras-related protein Rab11C)基因。方法利用PCR技术扩增阴道毛滴虫TvRab11C基因,与原核表达载体pET-28a连接,构建重组原核表达质粒pET-28a-TvRab11C,转化大肠杆菌BL21(DE3),IPTG诱导表达,SDS-PAGE分析表达产物的可溶性,Western blot分析表达产物的反应原性。结果重组原核表达质粒pET-28a-TvRab11C经双酶切及测序证明构建正确;表达的重组蛋白相对分子质量约为30 000,主要以包涵体形式存在,可被抗阴道毛滴虫多克隆抗体识别。结论成功克隆了阴道毛滴虫TvRab11C基因,并在E.coli BL21(DE3)中获得了表达,为进一步研究TvRas基因和G蛋白与阴道毛滴虫寄生能力和致病性的关系奠定了基础。Objective To clone the TvRabllC gene of Trichomonas vaginalis and express in prokaryotic cells. Methods The TvRabl1C gene was amplified by PCR from T. vaginalis and inserted into prokaryotic expression vector pET-28a. The construct- ed recombinant plasmid pET-28a-TvRabllC was transformed to E. coli BL21 （DE3） and induced by IPTG. The expressed product was analyzed for solubility by SDS-PAGE and for reactogenicity by Western blot. Results Both restriction analysis and sequencing proved that recombinant plasmid pET-28a-TvRabllC was constructed correctly. The expressed recombinant protein, with a relative molecular mass of about 30 000, mainly existed in a form of inclusion body, and was recognized by polyclonal antibody against T. vaginalis. Conclusion The TvRas gene of T. vaginalis was successfully cloned and expressed in E. coli BL21 （DE3）, which laid a foundation of further study on relationship of TvRas gene and protein to the parasitic ability and pathogenicity of T.vaginatis.

关 键 词：毛滴虫 阴道 TvRab11C基因 克隆 原核表达 

分 类 号：R382.21[医药卫生—医学寄生虫学] Q786[医药卫生—基础医学]