【摘要】 以pS189、pSV₂gpt、pMCi5为材料,构建了以EB病毒为基础,以酪氨酸tRNA琥珀突变校正基因SupF为靶基因,以黄嘌呤-鸟嘌呤磷酸核糖转移酶基因Eco gpt为选择标记基因的穿梭质粒pZH32。pZH32长9.4kb,克服了以SV40为基础的穿梭质粒自发突变率高以及宿主范围局限的缺点。SupF基因遗传背景清楚,长度126bp,便于进行序列分析。gpt基因使转化克隆的筛选更加方便。有利于建立一种化学诱变剂的分子检测系统,研究基因突变的类型和突变机制。更多还原

【Abstract】 The shuttle vector can replicate in both bacteria and eukaryotic cells. We have constructed a shuttle vector pZH32 based on the EB virus. Three component parts of the plasmid come from pSV2gpt, pS189 and pMCi5. An E. coli tyrosine suppressor tRNA gene, supF, was used as a mutagenic target in this plasmid, and a gpt gene was used as a selectable marker gene for transformed cells. This pZH32 based on EBV replicates autonomously in the nuclei of human cells. It hasa low background mutation frequency. The plasmid can be used to examine the mutagenic mechanism of potential mutagens and carcinogens in mammalian cells.更多还原