Cotransfer of syntenic human genes into mouse cells using isolated metaphase chromosomes or cellular DNA.
Chromosome-mediated gene transfer (CMGT) of the human genes for hypoxanthine phosphoribosyl transferase (HPRT) and cytosol thymidine kinase (TK1) into HPRT deficient mouse A9 cells or TK deficient Swiss mouse 3T3TK- cells was found to occur at frequencies at least one order of magnitude higher than DNA-mediated gene transfer (DMGT). The frequency of CMGT into 3T3TK- cells was reduced by more than an order of magnitude by a posttreatment of the recipient cells with dimethyl sulphoxide (DMSO). After CMGT, expression of the non-selected genes coding for galactokinase (GALK) and acid alpha-glucosidase (GAA), both syntenic with TK1, was observed in a number of transformants. From the pattern of cotransfer, a tentative gene ordering of CENTROMERE-GALK-TK1-GAA on human chromosome 17 was deduced. Chromosome-mediated cotransfer of X-linked human phosphoglycerate kinase (PGK) with HPRT was observed in two out of 33 A9 transformants analysed. DNA-mediated cotransfer of a syntenic gene was only observed for GALK, cotransferred with TK1 in two out of 18 TK+ transformants of mouse LTK- cells. Therefore, with murine cells as recipients of human donor genetic material, CMGT results in a higher frequency of transfer and a higher incidence of cotransfer of syntenic genes than DMGT using cellular DNA in the same cell system.
D000818:Animals; D002874:Chromosome Mapping; D002902:Chromosomes, Human, 16-18 / Q000648:ultrastructure*; D004247:DNA / Q000235:genetics; D004274:DNA, Recombinant; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005686:Galactokinase / Q000235:genetics*; D005786:Gene Expression Regulation; D005796:Genes; D008040:Genetic Linkage; D005959:Glucosidases / Q000235:genetics*; D006801:Humans; D006822:Hybrid Cells; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008677:Metaphase; D051379:Mice; D013937:Thymidine Kinase / Q000235:genetics*; D014170:Transformation, Genetic; D000520:alpha-Glucosidases / Q000235:genetics*
D004274:DNA, Recombinant; D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase; D013937:Thymidine Kinase; D005686:Galactokinase; D005959:Glucosidases; D000520:alpha-Glucosidases; D004121:Dimethyl Sulfoxide