DNA-mediated cotransfer of unlinked mammalian cell markers into mouse L cells.
Purified DNA from three different types of mammalian cells was precipitated with calcium phosphate and added to mouse L cells deficient in thymidine kinase (TK). Donor DNA was prepared from three cell lines: (a) mouse cells transfected with UV-inactivated herpes simplex virus (HSV) type 1, or a purified fragment of HSV carrying the TK gene (b) human HeLa cells, and (c( CHO, a cell line derived from Chinese hamster ovaries. Several hypoxanthine-aminopterin-thymidine resistant colonies were isolated from each experiment. The origin of the TK that is expressed in these cells was studied by polyacrylamide gel electrohporesis, isoelectric focusing, or heat stability. The TK in all instances was of the donor origin. To determine the extent of gene transfer we have assayed the CHO and HeLa DNA transfectants for galactokinase (GALK), a marker closely linked to TK, and 25 other isozymes representing a large number of different chromosomes. No cotransfer of GALK was observed, indicating that the size of the transferred DNA segment is limited. We observed that, in one instance, esterase-D, an unlinked marker of Chinese hamster origin, was transferred along with TK. These experiments indicate that nonselected markers can be transferred by this method, although at a low efficiency.
D000818:Animals; D006224:Cricetinae; D004247:DNA / Q000235:genetics*; D004950:Esterases / Q000235:genetics; D005686:Galactokinase / Q000235:genetics; D005796:Genes; D005818:Genetic Engineering / Q000379:methods*; D008040:Genetic Linkage; D006801:Humans; D007739:L Cells / Q000502:physiology*; D051379:Mice; D013937:Thymidine Kinase / Q000235:genetics*; D014162:Transfection; D014170:Transformation, Genetic
D004247:DNA; D013937:Thymidine Kinase; D005686:Galactokinase; D004950:Esterases