Hsu C, Kavathas P, Herzenberg L A
Nature. 1984;312(5989):68-9. doi: 10.1038/312068a0.
We have shown previously that transfection of mouse L-cells with DNA from JM, a human T-cell line expressing certain T-cell differentiation antigens, yields stable transfectants expressing one or another of these antigens. The identities of the antigens were confirmed by immunoprecipitation and SDS-polyacrylamide gel electrophoresis. We now report that our procedure--co-transfection with the chicken thymidine kinase gene (tk) and whole cellular DNA, selection with hypoxanthine-aminopterin-thymidine (HAT), and staining of the cells with fluorochrome-conjugated monoclonal antibodies and fluorescence-activated cell-sorter (FACS) selection--yields transfectants expressing a variety of cell-surface molecules (19 of 21 investigated), most at a frequency of about one per 10(3) Tk+ transformants. Of these, 9 of 12 were transferred and expressed as readily using DNA from cells which did not express the cell-surface antigens as from tissues or cells that did express them.
我们之前已经表明，用来自JM（一种表达某些T细胞分化抗原的人T细胞系）的DNA转染小鼠L细胞，可产生表达这些抗原中的一种或另一种的稳定转染子。通过免疫沉淀和SDS-聚丙烯酰胺凝胶电泳确认了抗原的身份。我们现在报告，我们的方法——用鸡胸苷激酶基因（tk）和全细胞DNA共转染，用次黄嘌呤-氨基蝶呤-胸腺嘧啶核苷（HAT）进行选择，并用荧光染料偶联的单克隆抗体对细胞进行染色以及用荧光激活细胞分选仪（FACS）进行选择——可产生表达多种细胞表面分子的转染子（在所研究的21种中有19种），大多数的频率约为每10(3)个Tk+转化子中有一个。其中，12种中的9种使用不表达细胞表面抗原的细胞的DNA与使用表达这些抗原的组织或细胞的DNA一样容易转移和表达。
