【摘要】 目的　构建融合蛋白FADDdel GFP真核表达载体 pFADDdel GFP ,用该融合基因转染小鼠胰岛瘤细胞 (NIT) ,从而抑制死亡受体介导的细胞内凋亡信号传导 ,以探讨IDDM的发病机制及防治措施。方法　采用基因重组技术将FADDdel定向连接于真核表达载体 pEGFP N1 ,用脂质体将重组子pFADDdel GFP导入哺乳动物细胞NIT并检测其表达 ,采用FACS和MTT法检测特异性T细胞对转染重组子NIT引起的细胞毒效应。结果　转染重组子 pFADDdel GFP的NIT可见绿色荧光蛋白表达 ;pFADDdel GFP转染的NIT对特异性T细胞介导的细胞毒有明显抵抗。结论　成功构建 pFADDdel GFP融合基因并获稳定表达 ;pFADDdel GFP转染NIT可有效抑制死亡受体介导的细胞内凋亡信号传导。更多还原

【Abstract】 Objective To construct an eukaryotic gene expressing vector of fusional protein FADDdel GFP, to inhibit the death receptor mediated intracellular apoptosis signal transduction by modifying the target cells with pFADDdel GFP for exploring the mechanism and therapy of IDDM. Methods The target gene of FADDdel was inserted into the pEGFP N1 vector. After transfecting pFADDdel GFP into mammalian cells NIT(mouse insulinoma cells), cytotoxicity of NIT transfected with pFADDdel GFP mediated by special T cells was detected by FACS and MTT. Results GFP was expressed in NIT transfected with pFADDdel GFP; NIT modified by pFADDdel GFP could resist special T cells mediated cytotoxicity. Conclusion We constructed pFADDdel GFP fusion gene and acquired stably expression. NIT modified by pFADDdel GFP can effectively inhibit intracellular apoptosis signal transducted by death receptor.更多还原