CAJ | 학술논문

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pLK01．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 pCDNA31．／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。
목적：포장표체TIPE2-shRNA적만병독과립。방법사순병합성7대침대TIPE2적shRNA서렬，장기극륭입pLK01．재체，경매절급측서정학후，장구건호적7충pLKO 1．-TIPE2-shRNA질립분별여 pCDNA31．／3× FLAG-TIPE2공전염293T세포，Western blot감정간우효솔병진행후속적병독포장。결과4호질립침묵효과최호，장기여 Non-target-shRNA질립분별진행병독포장，Western blot화면역형광증실병독구유량호적감염효솔화침묵효과。결론 TIPE2-shRNA적만병독재체구건급병독포장성공。
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 .