摘要
目的克隆表达细粒棘球绦虫亲环蛋白(EgCyP)基因,并对其进行生物信息学分析。方法从细粒棘球绦虫cD NA中扩增目的基因,克隆入表达载体pET28a,转化大肠埃希菌(E.coli)BL21(DE3),经异丙基βD硫代半乳糖苷诱导表达后,进行SDS PAGE和免疫印迹试验鉴定,并对其进行生物信息学分析。结果重组质粒pET28a EgCyP构建成功。SDS PAGE和免疫印迹试验结果显示,重组蛋白在E.coliBL21(DE3)中获得高效表达,重组蛋白EgCyP分子量约为22 kDa,可被细粒棘球绦虫感染犬血清识别。生物信息学分析显示该蛋白具有7个潜在的抗原表位。结论成功克隆出细粒棘球绦虫EgCyP基因并在E.coliBL21(DE3)中表达,为进一步研究其免疫原性奠定了基础。
Objective To clone and express EgCyP gene of Echinococcus granulosus and analyze EgCyP using bioinformatics. Methods Total RNAS of adult E. granulosus was extracted and reversedly transcripted to cDNA. EgCyP gene was amplified from cDNA and inserted into vector pET28a. Recombinant plasmid pET28a-EgCyP was transformed into E. coli BL21 （DE3） for expres- sion under the induction of IPTG. The expressed product was identified by SDS-PAGE and Western blotting. EgCyP was analyzed by the bioinformatics software. Results The EgCyP gene was successfully amplified from cDNA of adult E. granulosus and a fu- sion protein was expressed in E .coli BL21 （DE3）. The molecular weight of the expressed protein was about 22 kDa. The Western blotting indicated that the antigenicity of the protein was specific. The bioinformatics analysis revealed that there were 7 antigen ep- itopes in EgCyP. Conclusion EgCyP ofE. granulosus is cloned and expressed in E. coli BL21 （DE3） successfully, which might be the foundation for the further study of its immunogenicity.
出处
《中国血吸虫病防治杂志》
CAS
CSCD
2012年第3期294-297,302,F0003,共6页
Chinese Journal of Schistosomiasis Control
基金
国家自然科学基金(30872212)
国家重大科技专项(2012ZX10004-201)
关键词
细粒棘球绦虫
亲环蛋白
克隆
表达
生物信息学分析
Echinococcus granulosus
Cyclophilin
Clone
Expression
Bioinformatics analysis
作者简介
王莹,女,博士研究生。研究方向:寄生虫分子免疫学;通信作者:曹建平,E-mail:caojp@yahoo.com。
