布鲁菌感染小鼠RAW264.7细胞对IRG1基因的表达调控作用  

Lentivirus-mediated RNA interference targeted inhibition expression of mice IRG1 gene and its role in Brucellainfection

作　　者：孙良文[1] 孙长江[1] 闫静[1] 张明亮[1] 冯新[1] 韩文瑜[1] 

机构地区：[1]吉林大学动物医学学院,吉林长春130062

出　　处：《中国兽医学报》2014年第2期239-242,247,共5页Chinese Journal of Veterinary Science

基　　金：国家转基因生物新品种培育重大专项资助项目(2009ZX08009-163B);国家自然科学基金-青年基金资助项目

摘　　要：针对小鼠RAW264.7细胞IRG1基因设计4个RNA干扰靶位,筛选出最佳干扰序列构建shRNA慢病毒载体质粒并包装获得慢病毒颗粒,进而经嘌呤霉素筛选获得稳转细胞系,实现IRG1基因在RAW264.7细胞基因表达的沉默。并通过布鲁菌16M株及M5株感染基因沉默细胞对IRG1基因在布鲁菌感染中的作用进行研究。结果表明,慢病毒介导的shRNA高效、稳定地沉默了IRG1基因的表达,布鲁菌侵染RAW264.7细胞后IRG1基因表达上调。本试验为IRG1基因及相关调控基因抗布鲁菌病作用研究奠定了基础。Four shRNAs targeting mice RAW264.7 cells IRG1 gene were designed to select the best shRNA sequence construct lentivirus vector plasmid and pack virus particles,and then obtain sta ble cell line by puro screen,realize silence of IRG1 gene expression in RAW264.7 cells. The role of IRG1 gene in Brucella infection were studied by the Brucella 16M strain and M5 strain infectio ning gene silence cells. The results showed that the lentivirus mediated shRNA efficiently and stably silenced expression of IRG1 gene,its expression was up-regulated after Brucella infection RAW264.7 cells. This study lays the foundation for IRG1 and related regulating genes resistant Brucella research.

关 键 词：小鼠 RAW264 7细胞 RNA干扰 1RGl基因 布鲁菌 

分 类 号：S855.12[农业科学—临床兽医学]