首都医科大学学报 ›› 2006, Vol. 27 ›› Issue (2): 162-165.

• 专题报道 • 上一篇    下一篇

结核分枝杆菌黏附素HBHA的克隆表达及其免疫原性研究

李传友, 张旭霞, 张健源, 李卫民, 田苗, 邱云青, 孙照刚, 李妍, 端木宏谨   

  1. 北京市结核病胸部肿瘤研究所细菌免疫室
  • 收稿日期:2006-02-28 修回日期:1900-01-01 出版日期:2006-04-24 发布日期:2006-04-24

Cloning and Expression of Heparin-binding Haemagglutinin Adhesin of Mycobacterium Tuberculosis and Its Immungenicity

Li Chuanyou, Zhang Xuxia, Zhang Jianyuan, Li Weimin, Tian Miao, Qiu Yunqing, Sun Zhaogang, Li Yan, Duanmu Hongjin   

  1. Department of Bacteriology and Immunology, Beijing TB & Thoracic Tumor Research Institute
  • Received:2006-02-28 Revised:1900-01-01 Online:2006-04-24 Published:2006-04-24

摘要: 目的 应用克隆表达技术从大肠杆菌中获得重组的结核分枝杆菌黏附素HBHA蛋白,并以HBHA免疫小鼠观察抗HBHA抗体的产生并探讨其免疫原性.方法 1)应用PCR技术扩增结核分枝杆菌hbhA编码基因的特异片段.2)结核分枝杆菌hbhA编码基因的克隆及在大肠杆菌中的表达.3)应用克隆表达的HBHA蛋白免疫小鼠,并用ELISA法测定小鼠血清抗HBHA抗体水平.结果 1)结核分枝杆菌hbhA基因成功地克隆到PET-32 a(+)表达载体中,并在大肠杆菌中获得了高效表达.重组HBHA主要存在于包涵体中,纯化包涵体并获得了大量的重组HBHA.2)重组HBHA免疫小鼠可产生较高水平的抗HBHA抗体(>1:102 400),表明重组HBHA在纯化过程中仍保留较强的免疫原性.结论 结核分枝杆菌黏附素HBHA具有较好的免疫原性.

关键词: 分枝杆菌, 结核, 黏附素, 表达, 免疫原性

Abstract: Objective To clone and express hbhA gene of M.tuberculosis,and observe production of anti-HBHA antibody and its immunogenicity from mouse serum.Methods 1) The hbhA coding gene of M.tuberculosis were amplified by PCR method.2) hbhA gene was cloned into E.coli for expression of specific protein.3) HBHA was injected into mouse and observed production of anti-HBHA antibody by ELISA method.Results 1) The hbhA gene of M.tuberculosis could be cloned into PET-32a(+) expression vector,and got a high expression.2) The recombinant HBHA increased antibody titer more than 1000000 times of immunized mouse compared with control group.Conclusion HBHA of M.tuberculosis has good immunogenicity.

Key words: Mycobacterium, tuberculosis, adhesin, expression, immunogenicity

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