钙整合素结合蛋白1慢病毒干扰载体的构建及表达  

Construction and Expression of a Lentiviral Vector for RNA Interference of Calcium-and Integrin-binding Protein 1 Gene

作　　者：惠玲[1] 李晓云[1] 王晓辉[1] 王美亮[1] 杨霄鹏[1] 马明仁[1] 桑春艳[1] 王剑锋[1] 

机构地区：[1]兰州军区兰州总医院医学实验中心,甘肃省干细胞与基因药物重点实验室,兰州730050

出　　处：《解放军医药杂志》2014年第7期48-51,共4页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：国家自然科学基金(81372177);甘肃省自然科学基金(1107RJZA106);全军医药卫生科研基金目(CLZ12JA08)

摘　　要：目的构建人钙整合素结合蛋白1(calcium-and integrin-binding protein 1,CIB1)基因RNA干扰慢病毒载体,获得稳定下调CIB1表达的慢病毒。方法根据CIB1的序列,设计siRNA序列,克隆入慢病毒核心质粒pGV112中,并与辅助质粒一起转染293T细胞进行病毒包装,用慢病毒感染293T细胞并测定病毒滴度。以CIB1干扰慢病毒感染人胶质瘤细胞SHG44后,应用Western blotting方法检测CIB1的表达。结果成功构建CIB1干扰慢病毒载体,包装后获得慢病毒颗粒,病毒滴度检测可达6×108pfu/ml,感染细胞CIB1蛋白表达水平显著下调(P<0.01)。结论成功构建人CIB1基因RNA干扰慢病毒,为后续CIB1生物学功能研究奠定基础。Objective To construct a method of lentiviral vector for RNA interference （RNAi） of human calci-um- and integrin-binding protein 1 （CIB1） gene to obtain lentiviral stably down-regulated CIB1 expression. Methods The sequence of siRNA for CIB1 interference was cloned into the pGV112 vector, and viruses packaging with assistant plasmids in 293T cells was performed, and then 293T cells were infected with the lentivirus, the virus titer was detected. The CIB1 expression was detected by Western blotting after human glioma cells SHG44 were infected by CIB1 interfered in lentivirus. Results The lentiviral particle was packaged with a virus titer reaching 6 ＆#215; 108 pfu/ ml after interference vector CIB1 was successfully constructed, and the level of CIB1 protein expression was significantly down-regulated （P 〈0. 01）. Conclusion The lentiviral vector for RNA interference of CIB1 has been successfully constructed, and it pro-vides a foundation for further research of CIB1 biological function.

关 键 词：钙整合素结合蛋白1 慢病毒 胶质瘤 RNA干扰 

分 类 号：R34-33[医药卫生—基础医学]