Electric field mediated transformation: isolation and characterization of a TK+ subclone.
Transformation of mammalian TK- cells by a plasmid carrying the TK gene from Herpes virus simplex 1 (pAGO) was mediated by electroporation. The cells were treated either in suspension or growing in monolayers directly in the petri dish. The yield of transformation was between 8.10(-5) and 2.10(-4) per microgram DNA depending on the experimental conditions. The structure of the integrated DNA was investigated proving the occurrence of a duplication process that affected preferentially the pBR322 part of the pAGO DNA (60 copies per cell). The TK gene that gave the TK+ phenotype to the selected clone was present in less than 6 copies.
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D000818:Animals; D002460:Cell Line; D002469:Cell Separation; D002472:Cell Transformation, Viral; D006224:Cricetinae; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004279:DNA, Viral / Q000032:analysis; D015245:Deoxyribonuclease BamHI; D015246:Deoxyribonuclease EcoRI; D015252:Deoxyribonucleases, Type II Site-Specific; D004558:Electric Stimulation; D005260:Female; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D018139:Simplexvirus / Q000235:genetics*; D013937:Thymidine Kinase / Q000235:genetics*; D014162:Transfection
D004279:DNA, Viral; D013937:Thymidine Kinase; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI; D015246:Deoxyribonuclease EcoRI; C025630:CAGCTG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific