布鲁氏菌Ⅳ型分泌系统效应子VceC功能的初步研究  

Preliminary study on the function of brucella type Ⅳ secretion system(T4SS) effector VceC

作　　者：张沾[1] 陈创夫[2] 张辉[2] 张艳[2] 张娜[1] 李臻[1] 陈瑞花[2] 张豫[2] 

机构地区：[1]石河子大学生命科学学院,新疆石河子832003 [2]石河子大学动物科技学院,新疆石河子832003

出　　处：《中国预防兽医学报》2011年第9期685-688,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：973计划项目(2010CB30203);国家自然科学基金(31001046/30960288);高层次人才科研启动资金(RCZX200914)

摘　　要：为研究布鲁氏菌(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。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-N1 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.

关 键 词：羊种布鲁氏菌 Ⅳ型分泌系统 VceC 

分 类 号：S852.61[农业科学—基础兽医学]