[Genetic transformation of somatic cells. I. Clone of Chinese hamster cells defective in thymidine kinase and characterized by high transformation efficiency].
A characteristics is given of clone A238 of the Chinese hamster cells deficient in thymidine kinase (TK). The isolation procedure is described. Upon transformation with the aid of DNA of plasmids, containing thymidine kinase gene (tk-gene) of Herpes simplex virus type 1 (HSV1) clone A238 cells show frequency (7.10(-5) and efficiency (130 TK+ colonies per 1 microgram of plasmid DNA) compatible with those of mouse line LMtk- cells. Modified transformation and selection conditions of clone A238 cells expressing TK-gene of HSV1 are demonstrated. A simple method is described for discriminating somatic cells, expressing either their proper or a virus TK-gene according to the cloning efficiency of cells on the HAT medium containing thymidine in concentration 100 micrograms/ml. It is shown that at the fixed total DNA concentrations a complete replacement of the eukaryotic carrier DNA for the plasmid DNA, containing no tg gene of HSV1, decreases but only insignificantly the frequency and efficiency of transformation.
Glebov
O K
OK
Abramian
D S
DS
Tomilin
N V
NV
D000818:Animals; D002469:Cell Separation; D002472:Cell Transformation, Viral; D002999:Clone Cells / Q000201:enzymology; D006224:Cricetinae; D003412:Cricetulus; D004279:DNA, Viral / Q000235:genetics; D004526:Efficiency / Q000502:physiology; D005786:Gene Expression Regulation; D005814:Genes, Viral; D010957:Plasmids; D018139:Simplexvirus / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics*; D014170:Transformation, Genetic
D004279:DNA, Viral; D013937:Thymidine Kinase
Geneticheskaia transformatsiia somaticheskikh kletok. I. Klon kletok kitaĭskogo khomiachka, defektnykh po timidinkinaze, otlichaiushchiĭsia vysokoĭ éffektivnost'iu transformatsii.