人支架蛋白IQGAP1-shRNA慢病毒载体的构建及鉴定  

Construction and identification of RNA interference lentiviral vector for IQGAP1 gene

作　　者：张芮 王一兵[2] 林莉[2] 王法刚[2] 李强[2] 曹永倩[2] 

机构地区：[1]山东医学高等专科学校病理教研室,山东济南250002 [2]山东大学附属省立医院烧伤整形美容外科,山东济南250021

出　　处：《泰山医学院学报》2013年第8期561-564,共4页Journal of Taishan Medical College

基　　金：山东省自然科学基金(2006ZRB14130);山东省优秀中青年科学家科研奖励基金(BS2011SW049)

摘　　要：目的制备携带人支架蛋白IQGAP1基因小分子干扰RNA的重组慢病毒。方法分别设计合成针对IQGAP1的4个shRNA慢病毒干扰载体;分别转染293T细胞,利用Western blot验证shRNA的沉默效果,选择其中一个沉默效果较好的shRNA慢病毒干扰载体同源重组产生Lentivirus-IQGAP1-shRNA并测定病毒滴度。结果测序证实,构建携带IQGAP1-shRNA的慢病毒载体具有较好的沉默靶基因的效果,包装慢病毒,病毒滴度为2.0×l010TU/ml。结论成功制备携带IQGAP1-shRNA的重组慢病毒载体并包装出具高效感染力的慢病毒颗粒。Objective： To construct recombinant lentivirus with IQGAPl-short hairpin RNA （shRNA）. Methods： Four different kinds of lentivirns vectors containing IQGAPI-shRNA were designed and synthesized. 293T ceils were transfected by these IQGAPI-shRNAs, and the method of western blot was applied to verify the silence effects. One of four IQGAPI-shRNAs which had the best silence effect was chosen to recombine lentivirus vector and the titer of purified virus was deter- mined. Results： The titer of virus was 2.0×l010TU/ml. Conclusion： The lentivirus-IQGAPl-shRNA is constructed suc- cessfully.

关 键 词：支架蛋白1 RNA干扰 慢病毒载体 

分 类 号：R346[医药卫生—基础医学]