[细粒棘球绦虫亲环蛋白的克隆、表达及生物信息学分析]
[Cloning, expression and bioinformatics analysis of cyclophilin of Echinococcus granulosus].
作者信息
Wang Ying, Shen Yu-Juan, Yuan Zhong-Ying, Xu Yu-Xin, Han Xiu-Min, Zhou He-Jun, Wang Jun-Yun, Wang Hu, Cao Jian-Ping
机构信息
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; Laboratory of Parasite and Vector Biology, Ministry of Health, China.
出版信息
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi. 2012 Jun;24(3):294-7, 302.
OBJECTIVE
To clone and express EgCyP gene of Echinococcus granulosas 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 expression 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 fusion 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 epitopes in EgCyP.
CONCLUSION
EgCyP of E. granulosus is cloned and expressed in E. coli BL21 (DE3) successfully, which might be the foundation for the further study of its immunogenicity.
目的
克隆并表达细粒棘球绦虫的EgCyP基因，并用生物信息学方法对其进行分析。
方法
提取细粒棘球绦虫成虫的总RNA并反转录为cDNA。从cDNA中扩增EgCyP基因并将其插入载体pET28a。将重组质粒pET28a-EgCyP转化至大肠杆菌BL21(DE3)中，在IPTG诱导下进行表达。通过SDS-PAGE和Western印迹法对表达产物进行鉴定。用生物信息学软件对EgCyP进行分析。
结果
成功从细粒棘球绦虫成虫cDNA中扩增出EgCyP基因，并在大肠杆菌BL21(DE3)中表达出融合蛋白。表达蛋白的分子量约为22 kDa。Western印迹表明该蛋白具有特异性抗原性。生物信息学分析显示EgCyP中有7个抗原表位。
结论
细粒棘球绦虫的EgCyP在大肠杆菌BL21(DE3)中成功克隆并表达，这可能为进一步研究其免疫原性奠定基础。
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