含I-A^dα和β链cDNA的三顺反子逆转录病毒载体构建及真核表达  

Construction of Tricistronic Retroviral Vector Plasmid Containing both I-A^dα and I-A^dβ cDNA and its Expression in Eukeryotic Cells

作　　者：李煜[1] 钱书兵[1] 陈诗书[1] 钱关祥[1] 

机构地区：[1]上海第二医科大学人类基因治疗研究中心,上海200025

出　　处：《上海免疫学杂志》2001年第3期144-147,153,共5页Shanghai Journal of Immunology

摘　　要：为了协同表达MHCII类分子的两个链α和 β ,利用脑心肌炎病毒和脊髓灰质炎病毒的内核糖体进入位点 ,构建了含MHCII类分子α和 β链及选择标记新霉素磷酸转移酶基因的三顺反子逆转录病毒载体。经包装细胞包装成重组逆转录病毒后 ,感染NIH3T3、MM45T Li、COS7细胞 ,经PCR、RT PCR、Southern印迹、Northern印迹及流式细胞术在多水平上证实了α链和 β链的基因表达和MHCII类分子的正确组装。In order to express the two subunits of MHC class II molecule IA genes, the internal ribosome entry site(IRES) from encephalomyocarditis and poliomyelitis viruses are used to construct tricistronic retroviral vector, which contains genes of neomycin phosphotransferase, I A dα and I A dβ cDNA The transduction of NIH3T3, MM45T Li and COS7 cells with recombinant retrovirus packaged with those vectors by packaging cells(PA317) allows coordinated expression of I A dα,I A dβ and neomycin phosphotransferase The correct expression and assembling have been confirmed by PCR, RT PCR, Southern blot , Northern blot and flow cytometry

关 键 词：MHC Ⅱ类分子 三顺反子逆转录病毒载体 内核糖体进入位点 基因治疗 肿瘤 

分 类 号：R730.54[医药卫生—肿瘤]