期刊论文

Wu, Y.-M.

中文

中华微生物学和免疫学杂志

0254-5101

2010

30

5

487-492

10.3760/cma.j.isan.0254-5101.2010.05.022

国家自然科学基金（30870134）；

本校为第一且通讯机构

目的研究肺炎嗜衣原体(Chlamydophila pneumoniae, Cpn)衣原体蛋白酶样活性因子 (Chlamydial protease-like activity factor,CPAF)能否在体外诱导人单核细胞THP-1产生前炎症细胞因子和凋亡,为进一步探索Cpn,感染宿主致病的分子机制提供实验依据.方法将Cpn,CPAF全基因克隆于pGEX6p-2载体上,在大肠杆菌(E.coli)中诱导表达重组蛋白,经去内毒素纯化柱和琼脂糖凝胶FF获得纯化且无脂多糖(LPS)污染的重组蛋白GST-CPAF.以不同浓度的该蛋白作用于THP-1, ELISA检测TNF-α和IL-6水平.MTT法检测经该蛋白处理后THP-1的增生或抑制作用,用 Hoechst33258荧光染色、DNA片段化分析、Annexin V-FITC-PI染色法检测细...

Objective: To express and purify Chlamydial protease-like activity factor (CPAF) from Chlamydophila pneumoniae, for investigating the effect of its recombinant protein GST-CPAF in inducing human monocytic cells to secrete proinflammatory cytokines and cell apoptosis. Methods: The recombination expression plasmid pGEX6p-2/CPAF from Chlamydophila pneumoniae was transformed into E. coli. The recombination GST-CPAF was expressed after induction by IPTG, and purified by a agarose gel FF. Human monocytic cells were stimulated by the GST-CPAF to test ...