布鲁氏茵Ⅳ型分泌系统效应子VceC功能的初步研究

摘要

To explore the biological function of VceC as an effector of brucella type IV secretory system, the vceC gene was amplified from B. Melitensis 16M strain by PCR and cloned into the eukaryotic expression vector pEGFP-Nl to construct recombinant plasmid pEGFP-vceC. The recombinant plasmid was transfected into HPT-8 cells by liposome and the expression of GFP-vceC fusion protein was identified by western blot and fluorescence microscopy. The test results also demonstrated that the cell apoptosis, the NO release and the LDH activity were increased in HPT-8 cells transfected with pEGFP-vceC, comparing with control cells (p<0.05). This study provided preliminary evidence that the VceC had cytotoxic effect to HPT-8 cells.%为研究布鲁氏茵(Brucella)Ⅳ型分泌系统效应子VceC的功能,本研究从羊种布鲁氏菌16M株基因组中克隆vceC基因片段,插入pEGFP-N1中,构建真核表达重组pEGFP-vceC.经脂质体转染HPT-8细胞,荧光显微镜观察、western blot鉴定目的蛋白在HPT-8细胞中表达情况,检测细胞凋亡、NO释放量及LDH活力.Western blot检测显示VceC在细胞中表达,转染pEGFP-vceC后的细胞凋亡、细胞上清液中NO释放量及LDH活力比对照组显著增加(p＜0.05).本研究初步表明VceC蛋白具有细胞毒性作用,为进一步研究VceC的功能奠定了基础.