布鲁氏菌介导对细胞类泛素SUMO1表达影响的研究  

Effection on expression of SUMO1 in cells infected by Brucella

作　　者：监通[1] 陈创夫[1] 张辉[1] 张俊波[1] 张科[1] 李跃峰[1] 李蕊[1] 王讲德[1] 程婷婷[1] 

机构地区：[1]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》2013年第12期1012-1015,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家973项目(2010CB30203);国家自然基金(31001046;31260596);石河子大学杰出青年项目(2012ZRKXJQ02)

摘　　要：为研究布鲁氏菌侵染细胞后对SUMO1表达的影响,本研究采用Ni柱亲和层析法纯化SUMO1蛋白,制备兔抗SUMO1多克隆抗体,布鲁氏菌M5-90和16M菌株分别侵染巨噬细胞后,通过免疫印迹方法和荧光实时定量方法分别检测细胞中类泛素SUMO1蛋白表达水平与mRNA转录水平。结果显示,表达并纯化了鼠源类泛素SUMO1蛋白,并制备了兔抗SUMO1多克隆抗体;布鲁氏菌M5-90和16M菌株侵染巨噬细胞后,SUMO1蛋白表达水平与mRNA转录水平在12 h内均呈现先上升后下降的趋势,M5-90菌株侵染4 h时SUMO1表达量最高(p<0.01),16M菌株侵染8 h时SUMO1表达量最高(p<0.01)。实验结果表明布鲁氏菌在侵染细胞时能够引起类泛素SUMO1表达的增强,对布鲁氏菌在宿主细胞中存活和细胞抗菌均起一定作用。To study the expression of SUMO1 in macrophage infected with Brucella M5-90 or 16M strain, the mRNA levels of SUMO1 were detected by real-time RT-PCR and the SUMO1 expressions were assayed by western blot with the polyclonal antibodies against SUMO1 which was prepared in rabbit immunized with the purified prokariotic expressed SUMO1. The results shown that the levels of SUMO1 protein expression and mRNA transcription emerged that both ascended at the early stage and then declined during 12 hours after macrophage infected with M5-90 and 16M strains, and SUMO1 expression levels in macrophage infected with M5-90 and 16M strain reached peak level at 4 hours （P〈0.01） and 8 hours （p〈0.01）, respectively. The expression of SUMO1 was able to lead to ascend by Brucella infecting, which indicated that SUMO1 play important roles in Brucella survival and anti-microbial during the process of Brucella infecting macrophage.

关 键 词：多克隆抗体 布鲁氏菌 SUMO1 

分 类 号：S852.61[农业科学—基础兽医学]