中南医学科学杂志Issue(3)：229-232,4.
鹦鹉热嗜衣原体CPSIT_0018融合蛋白原核表达载体的构建、表达及其在Hela细胞中的定位
Construction and Expression of the Prokaryotic Clone of Recombinant Protein CPSIT_0018 and Localization in Hela Cells
摘要
Abstract
Objective The aim is to construct the prokaryotic expression plasmid of CPSIT_0018 gene from Cps 6BC strain;to express and purify the recombinant protein His-CPSIT_0018;and to localize the endogenous CPSIT_0018 pro-tein in Cps-infected Hela cells. Methods CPSIT_0018 gene was cloned by PCR and inserted into the expression vector pET30a to construct pET30a-CPSIT_0018. The recombinant plasmid was transformed into E. coli BL21,and the recombinant protein was expressed and purified. The purified protein was used to immunize BALB/c mice to produce polyclonal antibod-y,which were subsequently used to localize the endogenous CPSIT_0018 protein by indirect immunofluorescence assay (IFA). Results The recombinant expression plasmid pET30a-CPSIT_0018 was successfully constructed,and then the recombinant protein was expressed and purified. IFA showed that the distribution pattern of the CPSIT_0018 protein was similar to that of the major outer membrane protein ( MOMP) ,but not to that of inclusion membrane protein A ( IncA) .Conclusion CPSIT_0018 protein was successfully expressed and purified in prokaryotic expression system;CPSIT_0018 is located on the bacterial organism in Cps-infected Hela cells.关键词
鹦鹉热嗜衣原体/CPSIT_0018/克隆表达/细胞定位Key words
Chlamydophila psittaci/CPSIT_0018/cloning and expression/cellular localization分类
医药卫生引用本文复制引用
贺庆芝,曾怀才,陆春雪,胡艳群,陈芝喜,王川,吴移谋..鹦鹉热嗜衣原体CPSIT_0018融合蛋白原核表达载体的构建、表达及其在Hela细胞中的定位[J].中南医学科学杂志,2014,(3):229-232,4.基金项目
国家自然科学基金(81202323),湖南省科技厅项目(2012FJ6009),湖南省卫生厅项目(B2012-042),南华大学研究生科研创新项目(2012XCX13) （81202323）
