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.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
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)
作者简介
张沾（1985-），女，河南周口人，硕士研究生，研究方向为分子免疫学．
通信作者：E-mail：ccf-xb@163．com；
allanzhh@yahoo．com
