摘要
目的在大肠杆菌中表达肺炎衣原体（Cpn）热休克蛋白10（Hsp10），并观察该蛋白诱生巨噬细胞（Mφ）分泌TNF-α和IL-6等促炎因子的作用。方法将Hsp10基因克隆于pQE30载体，在大肠杆菌（E．coli）M15中表达重组蛋白rHsp10，经亲和层析获得纯化rHsp10。以不同浓度的rHsp10作用于小鼠MφRAW264．7，酶联免疫吸附试验（ELISA）检测细胞因子TNF-α和IL-6。结果成功构建了pQE30/Hsp10基因，并在大肠杆菌M15中得到高效表达，该蛋白能诱导小鼠Mφ以时间、剂量依赖方式分泌TNF-α和IL-6。结论表达并纯化的rHsp10蛋白能诱导Mφ分泌TNF-α和IL-6等促炎因子。
Objective To express and to purify Chlamydia pneumoniae （ C. pneumoniae） heat shock protein 10（Hsp10） and for investigating the effect of the protein in inducing proinflammatory cytokines secreted by routine macrophages. Methods Hsp 10 gene was cloned into prokaryotic expression vector pQE30. The recombination expression plasmid pQE30/Hsp10 was transformed into E. coli. The recombination Hsp10（rHsp10）, which had a 6-His tag, was expressed after induction by IFTG and purification. Murine macrophages were stimulated to test the production of tumor necrosis factor α（TNF-α） and interleukin-6（IL-6） by ELISA. Results The recombination plasmid pQE30/Hsp10 was successfully constructed with high level expression of rHsp10 in E. coli. The rHsp10 stimulated murine macrophages to produce proinflamatory cytokines including TNF-α and IL-6 in a dose- and time-dependent manner. Conclusion C. pneumoniae rHsp10 can induce the secretion of TNF-α and IL-6 by murine macrophages.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2007年第1期24-28,共5页
Chinese Journal of Microbiology and Immunology
基金
湖南省卫生厅资助重点课胚（A03-006）
关键词
肺炎衣原体
热休克蛋白10
促炎因子
巨噬细胞
Chlamydia pneumoniae
Heat shock protein 10
Proinflammatory cytokines
Macrophage
