中国强毒赖型钩端螺旋体新基因OmpL17的表达及其免疫原性  

Expression and Immunity Reaction of a Novel Gene OmpL17 of the Strong Virulent L. Interrogans Serovar Lai in China

作　　者：朱庆平[1] 赵计林[1] 鲍朗[1] 张会东[1] 赵明才[1] 黎光[1] 

机构地区：[1]四川大学华西基础医学与法医学院感染免疫研究室,成都610041

出　　处：《生物医学工程学杂志》2005年第2期250-253,共4页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目 (3 0 0 70 670)

摘　　要：将Omp L17基因克隆于p GEX- 1λT载体,在大肠杆菌JM10 9中表达分子量约为5 4 KDa的GST-Omp L17融合蛋白,凝血酶切割后得到了大小约2 8KDa的Omp L17蛋白。用纯化的Omp L17蛋白免疫大白兔,制备了高滴度的特异性抗体(1∶4 896 )。本研究获得了纯化的Omp L17及其特异性抗体,为该外膜蛋白致病作用及免疫保护力研究奠定基础。This study was conducted to potentiate the expression of outer membrane protein OmpL17 of the strong virulent L. interrogans serovar Lai and investigate its immunogenicity in rabbits. The OmpL17 was cloned into prokaryotic expression vector pGEX-1λT. The recombination expression plasmid pGEX-OmpL17 was transformed into E. Coli JM109. The GST fused protein GST-OmpL17 was expressed after induction by IPTG, then GST-tag was by thrombin and purified using Bulk GST purification Modules. SDS-PAGE and Western blotting analysis indicated that the molecular weight of GST-OmpL17 and OmpL17 was about 54KDa and 28KDa respectively. The outer membrane protein OmpL17 was subcutaneously injected into rabbits and high titre anti-OmpLl7 antibody was obtained （1 ： 4896） which could conjugate specifical with OmpL17. In conclusion, OmpL17 and specifical anti-OmpL17 antibody were obtained, which provided an experimental basis for researching pathogenic effect and immunity functions of OmpL17.

关 键 词：OmpL17 钩体外膜蛋白 GST融合表达 免疫原性 赖型钩端螺旋体 新基因 特异性抗体 中国 融合蛋白 JM109 

分 类 号：R392[医药卫生—免疫学]