Genetic linkage but independent expression of functional HSV-1 tk and mammalian aprt genes after cotransfer to L cells.
DNA-mediated gene transformation of mouse Ltk-aprt-hprt-cells was used to obtain stable, doubly selected transformants simultaneously expressing herpes virus thymidine kinase (TK) and mammalian adenine phosphoribosyltransferase (APRT). Cotransformants occurred at a frequency of 5 X 10(-6), a similar frequency for the transfer of the aprt marker has been previously observed. Isozyme and Southern blot analysis show that the TK and APRT expressed in these transformants resulted from gene transfer. For one stable cotransformant, [3H]thymidine [( 3H]TdR) selection against TK activity resulted in the loss of APRT activity as well, suggesting that these genes had become genetically linked together. Similarly selection against APRT expression resulted in the loss of a subset of the transferred herpes simplex virus tk genes. 5-Bromodeoxyuridine (BUdR) selected TK- variants differed from [3H]TdR selected TK- variants, in that they retained tk genes. However, BUdR-selected variants expressed full levels of APRT. Therefore, even though the transferred tk and aprt genes had become genetically linked together, they were, in this case, independently expressed since these cells were phenotypically TK- and APRT+.
Block
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Brzykcy
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Hastie
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Hughes
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D000228:Adenine Phosphoribosyltransferase / Q000235:genetics*; D000818:Animals; D001973:Bromodeoxyuridine; D002460:Cell Line; D004274:DNA, Recombinant / Q000378:metabolism; D005814:Genes, Viral; D008040:Genetic Linkage; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase; D007739:L Cells; D051379:Mice; D010430:Pentosyltransferases / Q000235:genetics*; D010641:Phenotype; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D013937:Thymidine Kinase / Q000235:genetics*; D014162:Transfection
D004274:DNA, Recombinant; D010430:Pentosyltransferases; D000228:Adenine Phosphoribosyltransferase; D007041:Hypoxanthine Phosphoribosyltransferase; D013937:Thymidine Kinase; D001973:Bromodeoxyuridine