[Genetic transformation of somatic cells. II. An analysis of the status of the plasmid nucleotide sequences in chromosomal DNA and the thymidine kinase activity in transformant clone cells].
Chinese hamster A238 TK- -cells were transformed with plasmids (derivatives of pBR325) containing thymidine kinase (TK) gene of Herpes simplex virus type 1 (HSV1). The results of dot- and blot-hybridization indicate the presence of pBR325 sequences in the chromosomal fractions of DNA in the transformant clones. These sequences are probably tandemly arranged, and each cluster contains 25--50 copies. SV40 sequences cloned in pBR325 were introduced into the Chinese hamster cells by co-transformation with TK-gene of HSV1-containing plasmid DNA, and all the co-transformant clones selected for TK+-phenotype were shown by hydridization to contain 3V40 DNA fragments. Isoelectrofocusing in polyacrylamide gel shows that thymidine kinase from TK+-transformant clones is of viral type (isoelectric point 7), in contrast to the cellular enzyme (coded by chromosomal gene) having alkaline isoelectric point (pH 9). The results suggest that the true TK+-transformant cells are selected by the procedure used in this study.
Tomilin
N V
NV
Podgornaia
O I
OI
Abramian
D S
DS
Glebov
O K
OK
D000818:Animals; D001483:Base Sequence; D002472:Cell Transformation, Viral; D002875:Chromosomes / Q000648:ultrastructure*; D002999:Clone Cells / Q000201:enzymology / Q000648:ultrastructure; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005814:Genes, Viral; D007525:Isoelectric Focusing / Q000379:methods; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics; D013937:Thymidine Kinase / Q000032:analysis / Q000235:genetics*; D014170:Transformation, Genetic
D004247:DNA; D013937:Thymidine Kinase
Geneticheskaia transformatsiia somaticheskikh keltok. II. Analiz sostoianiia plazmidnykh nukleotidnykh posledovatel'nosteĭ v khromosomnoĭ DNK i aktivnosti timidinkinazy v kletkakh transformantnykh klonov.