期刊论文

Chen, Y.Y.

中文

细胞与分子免疫学杂志

1007-8738

2011

27

9

954-958

10.13423/j.cnki.cjcmi.006131

国家自然科学基金资助项目（30870134）；

本校为第一机构

目的: 在E.coliBL21中表达肺炎嗜衣原体Cpn0810, 并研究该蛋白能否诱导人单核细胞(THP-1)产生TNF-α和IL-6等促炎因子和细胞凋亡。方法: PCR扩增Cpn0810蛋白编码基因, 构建pGEX6p-2/Cpn0810重组质粒, 在E.coliBL21中诱导表达, 经ToxinEraser纯化柱纯化后, 用不同浓度的GST-Cpn0810作用THP-1细胞, ELISA法检测TNF-α和IL-6的水平, Hoechst33258荧光染色、AnnexinV-FITC-PI染色法检测细胞凋亡情况。结果: 构建了重组质粒pGEX6p-2/Cpn0810, 并在E.coliBL21菌中高效表达。该蛋白能诱导THP-1细胞以时间、剂量依赖方式表达TNF-α和IL-6; 当10 mg/L GST-Cpn0810处理THP-1细胞24 h后, 形态学上, 细胞表现...

AIM: To expresse the Chlamydia pneumoniae Cpn0810 in E.coli BL21, and to study weather could it inducing proinflamatory cytokines including TNF-alpha and IL-6 in human monocytic (THP-1) and cell apoptosis. METHODS: Polymerase chain reaction(PCR) was used to amplify the Cpn0810 gene, PCR products were purified and cloned into the prokaryotic expression vector pGEX6p-2. The restriction plasmids pGEX6p-2/Cpn0810 confirmed by PCR and sequencing was transformed into E.coli BL21. The recombinant protein was purified with glutathione S-transferase (GST) resin chromatography of Novagen after renaturat...