靶向TIPE2基因的shRNA慢病毒载体构建及病毒包装  

Construction and package of recombinant lentivirus expressing shRNA targeting TIPE2 gene

作　　者：王耀辉[1] 王明丽[1] 陈九格[1] 张晶[1] 马远方[1] 

机构地区：[1]河南大学医学院,河南开封475004

出　　处：《河南大学学报（医学版）》2014年第2期120-122,134,共4页Journal of Henan University:Medical Science

基　　金：河南大学校内基金资助(2011YBZR005)

摘　　要：目的包装表达TIPE2-shRNA的慢病毒颗粒。方法查询并合成7对针对TIPE2的shRNA序列,将其克隆入pLK0.1载体,经酶切及测序正确后,将构建好的7种pLKO.1-TIPE2-shRNA质粒分别与pCDNA3.1/3×FLAG-TIPE2共转染293T细胞,Western blot鉴定干扰效率并进行后续的病毒包装。结果 4号质粒沉默效果最好,将其与Non-target-shRNA质粒分别进行病毒包装,Western blot和免疫荧光证实病毒具有良好的感染效率和沉默效果。结论 TIPE2-shRNA的慢病毒载体构建及病毒包装成功。Objective To package the recombinant lentivirus expressing TIPE2 shRNA .Methods Inquiring and synthesizing seven pairs shRNA sequences targeting TIPE2 ,then inserted into pLKO .1 lentivirus vector .After the constructions were verified to be correct ,co-transfected them with pCDNA3 .1/3 × FLAG TIPE2 plasmids into 293T cells , respectively . Then , Non target shRNA , the highest silence efficiency plasmid were co-transfected with psPAX2 , pMD2 .G into 293T cells for lentivirus packaging , respectively .Results The No .4 plasmid was screened by Western blotting that has the highest silence efficiency and followed by virus packing . The infection and silence efficiency was confirmed by immunofluorescence and Western blot . Conclusion The recombinant lentivirus expressing shRNA targeting TIPE2 gene was packaged successfully .

关 键 词：TIPE2 SHRNA 慢病毒包装 基因 

分 类 号：Q782[生物学—分子生物学]