免疫调节因子TIPE2慢病毒载体的构建及病毒包装  

Construction of human immune regulator TIPE2 lentiviral vector and lentivirus packaging

作　　者：王耀辉[1] 娄强[1] 陈九格[1] 张军[1] 马远方[1] 

机构地区：[1]河南大学医学院抗体药物河南省工程实验室,开封475004

出　　处：《中国免疫学杂志》2013年第10期1077-1079,共3页Chinese Journal of Immunology

基　　金：国家自然科学基金资助(30972687)

摘　　要：目的:TIPE2[Tumor necrosis factor(TNF)-αinduced protein 8-like 2(TNFAIP8L2),TIPE2]慢病毒载体的构建及病毒包装。方法:首先构建带有3×FLAG标签的pCDNA3.1-3×FLAG-TIPE2质粒,确认表达后再将标签连同TIPE2序列一起克隆入pCDH-CMV-MCS-EF1-Puro慢病毒表达载体,然后与pMD2.G、psPAX2共转染293T细胞并收获上清,进一步用含病毒颗粒的上清感染PLC/PRF/5及HEK293T细胞,Western blot检测感染后TIPE2蛋白表达情况。结果:pCDNA3.1-3×FLAGTIPE2及pCDH-CMV-MCS-EF1-Puro-TIPE2质粒构建成功,并能最终产出具有感染性的病毒颗粒。结论:成功构建了TIPE2慢病毒载体并获得具有感染性的病毒颗粒,为进一步研究TIPE2功能提供了有力的支持。Objective：To package lentivirus carrying human TIPE2 （Tumor necrosis factor （TNF）-ct induced protein 8-like 2 （TNFAIP8L2）, TIPE2） gene. Methods： The human TIPE2 gene was cloned into pDNA3. 1-3 x FLAG vector to generate pCDNA3.1- 3× FLAG-TIPE2 plasmid, after sequencing, 3× FLAG-TIPE2 fragment was inserted into pCDH-CMV-MCS-EF1-Puro lentiviral vector. The pCDH-CMV-MCS-EFI-Puro/3× FLAG-TIPE2 plasmids were co-transfected with pMD2. G,psPAX2 plasmids into HEK293T cells to harvest infectious virus particles, followed by infecting PLC/PRF/5 and HEK293T cells. Results： The TIPE2 expressing lentiviral vector was constructed successfully, and finally got infectious virus particles. Conclusion： The human TIPE2 recombinant lentiviral vector was successfully constructed and packaged.

关 键 词：免疫调节因子 TIPE2 慢病毒 病毒包装 

分 类 号：R392-3[医药卫生—免疫学]