中国免疫学杂志Issue(6)：790-794,5.DOI:10.3969/j.issn.1000-484X.2015.06.015
结核分枝杆菌Rv2460 c基因的克隆,表达及编码产物的免疫特性分析
Cloning and expression of Rv2460 c gene in mycobacterium tuberculosis and analysis of immune characteristics of its coding product
摘要
Abstract
Objective:To clone and express Mycobacterium tuberculosis(Mtb) Rv2460c gene(encoding ClpP2 protein),and evaluate the immunogenicity of its coding product. Methods: The recombinant plasmid of pET32a (+) vector-ClpP2 that H37Rv Rv2460c gene was cloned into the plasmid pET32a(+)vector,was transformed into E. coli BL21(DE3)and induced expression by IPTG,then purified by affinity chromatography. The recombinant protein was confirmed by SDS-PAGE and Western blot. The analysis of immunogenicity of Mtb ClpP2 and its epitope prediction were performed by bioinformatic methods. The antibody levels of polyclonal antibody titer against ClpP2 protein in rabbits and TB patients′ serum were detected by ELISA. Results: The recombinant ClpP2 protease was expressed as inclusion bodies in E. coli. The purity of purified protein was 93% by bandscan software analysis. The bioin-formatics analysis shows Mtb ClpP2 protein has multiple preponderant B cell and T cell epitopes. Rabbit antiserum titer was 1∶64 000;Serum anti-ClpP2 antibody levels in TB patients was higher than that in healthy control subjects. Conclusion:The recombinant ClpP2 protein was purified, and specific Rabbit anti-ClpP2 polyclonal antibody was prepared successfully. Experiment and bioinformatic information studies showed that Mtb ClpP2 protease has strong immunogenicity.关键词
结核分枝杆菌/重组蛋白ClpP2/免疫原性Key words
Mycobacterium tuberculosis/Recombinant ClpP2 protein/Immunogenicity引用本文复制引用
康月茜,张春燕,穆柳青,卢楠,杨春,李岱容..结核分枝杆菌Rv2460 c基因的克隆,表达及编码产物的免疫特性分析[J].中国免疫学杂志,2015,(6):790-794,5.基金项目
本文由国家青年科学基金( No．81101216)和国家临床重点专科专项经费资助(卫办医政函[2012]649号)资助。 （ No．81101216）
