Cell-surface antigens expressed on L-cells transfected with whole DNA from non-expressing and expressing cells.
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.
D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis / Q000235:genetics*; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D004247:DNA / Q000235:genetics*; D006680:HLA Antigens / Q000235:genetics; D006801:Humans; D007739:L Cells / Q000201:enzymology / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology; D051379:Mice; D013045:Species Specificity; D013601:T-Lymphocytes / Q000276:immunology; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D006680:HLA Antigens; D004247:DNA; D013937:Thymidine Kinase