论文部分内容阅读
在培养细胞中稳定表达麻疹病毒十分困难,这可能是由于病毒蛋白的细胞毒性所致。为了解决这一问题,用含有人类金属巯基组氨酸三甲基内盐Ⅱ基因可诱导启动子的HindⅢ至Xba Ⅰ片段取代含猿病毒40(SV40)早期启动子的pcD载体的HindⅢ至Xho Ⅰ区域,构建成pMT载体。将Xba Ⅰ位点转换为Xho Ⅰ位点。将含有麻疹病毒Edmonston株全长cDNA、或N基因、或M基因的质粒pcD-N 7或pcD-Mzi的Xho Ⅰ片段,插入pMT载体的Xho Ⅰ位点,分别产生pMT-N和pMT-M质粒。从pFR 400质粒中切下含DHFR＊基因蛋白密码区的Nco Ⅰ片段,
It is very difficult to stably express the measles virus in cultured cells, probably due to the cytotoxicity of the viral protein. To solve this problem, HindIII to XhoI of the pcD vector containing the simian virus 40 (SV40) early promoter was replaced with a HindIII to XbaI fragment containing the inducible promoter of human metal thiol histidine trimethylinosine II Region, construct into pMT vector. The Xba I site was converted to the Xho I site. The XhoI fragment of plasmid pcD-N7 or pcD-Mzi containing the full-length cDNA of the Edmonston strain of measles virus or the N gene or the M gene was inserted into the XhoI site of the pMT vector to generate pMT-N and pMT-M Plasmid. The Nco I fragment containing the codon for the DHFR * gene protein was excised from the pFR 400 plasmid,