[Genetic transformation of somatic cells. IX. The loss of the transformant phenotype is accompanied by rearrangements in the plasmid DNA containing the thymidine kinase gene of the herpes virus].
As demonstrated by dot-hybridization, the cells of HT-subclones isolated from the cells of transformant clones cultured on a non-selective medium differ significantly in the number of copies of thymidine kinase gene (tk-gene) of Herpes simplex virus (HSV1). Since the cells of transformant clones lose thymidine kinase-positive (TK+) phenotype during cultivation, this data are indicative of high frequence rearrangements in the region of transforming DNA as responsible for the transformant phenotype nonstability. These rearrangements, among other things, induce alterations in the number of copies of tk gene of HSV1. The analysis of cells of subclones isolated on a medium containing 5-bromodeoxyuridine (BrdU) shows that the number of copies of tk gene of HSV1 decreases as compared to the cells of parental clones. The decrease in the number of copies of tk gene of HSV1 in a row of BrdU-resistant subclones is accompanied by simultaneous increase in the number of sequences of pBR325 plasmide DNA to which tk gene of HSV1 is linked covalently in the pST826 plasmide introduced into cells of transformant clones. This evidence implies a most complex nature of transforming DNA rearrangements reducing the number of copies of tk gene of HSV1 due possibly to a genetic correction. The analysis of results permits a hypothesis that instability of cells in transformant phenotype may be determined by the genetic instability of insertion type. The rate of the loss of transformant phenotype depends on the frequency of rearrangements in the transforming DNA locus.
D000818:Animals; D002472:Cell Transformation, Viral / Q000187:drug effects; D002999:Clone Cells / Q000187:drug effects / Q000201:enzymology; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics*; D004351:Drug Resistance; D005814:Genes, Viral / Q000187:drug effects; D010641:Phenotype; D010957:Plasmids / Q000187:drug effects; D018139:Simplexvirus / Q000235:genetics*; D013937:Thymidine Kinase / Q000235:genetics*; D014170:Transformation, Genetic / Q000187:drug effects
D004279:DNA, Viral; D004247:DNA; D013937:Thymidine Kinase
Geneticheskaia transformatsiia somaticheskikh kletok. IX. Poteria transformantnogo fenotipa sopriazhena s perestroĭkami plazmidnoĭ DNK, soderzhashcheĭ gen timidinkinazy virusa gerpesa.