Genetic linkage but independent expression of functional HSV-1 tk and mammalian aprt genes after cotransfer to L cells

T Block, J Brzykcy, N Hastie, R G Hughes

Research output: Contribution to journalArticlepeer-review

Abstract

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+.

Original languageEnglish
Pages (from-to)311-6
Number of pages6
JournalCanadian Journal of Microbiology
Volume31
Issue number4
Publication statusPublished - Apr 1985

Keywords

  • Adenine Phosphoribosyltransferase
  • Animals
  • Bromodeoxyuridine
  • Cell Line
  • DNA, Recombinant
  • Genes, Viral
  • Genetic Linkage
  • Humans
  • Hypoxanthine Phosphoribosyltransferase
  • L Cells (Cell Line)
  • Mice
  • Pentosyltransferases
  • Phenotype
  • Simplexvirus
  • Thymidine Kinase
  • Transfection

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