Episkopou V, Murphy A J, Efstratiadis A
Proc Natl Acad Sci U S A. 1984 Aug;81(15):4657-61. doi: 10.1073/pnas.81.15.4657.
We have inserted into a retroviral vector the structural information encoding the selectable bacterial genes neo and gpt linked, respectively, to the promoters of the herpesvirus thymidine kinase gene and the rat preproinsulin II gene. We have used this recombinant retrovirus construction to transduce insulinoma cell lines that are positive or negative for insulin expression. Selection of the transductants for neo yielded resistant colonies from all of the cell lines, while selection for gpt yielded resistant transductants in relatively high frequency only from insulin-producing cells. The 5' end of the gpt transcripts maps to the authentic preproinsulin capping site of the construction.
我们已将编码可选择细菌基因neo和gpt的结构信息分别连接到疱疹病毒胸苷激酶基因启动子和大鼠前胰岛素原II基因启动子上,并插入逆转录病毒载体。我们利用这种重组逆转录病毒构建体转导胰岛素表达呈阳性或阴性的胰岛素瘤细胞系。对neo进行转导子选择时,所有细胞系均产生了抗性菌落,而对gpt进行选择时,仅在产生胰岛素的细胞中以相对较高的频率产生了抗性转导子。gpt转录本的5'端定位于构建体的真实前胰岛素原帽位点。