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PMID: 2990461 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Electric field mediated transformation: isolation and characterization of a TK+ subclone.

Biochemical and biophysical research communications ·Vol. 129 ·No. 3 ·1985-06-28 ·Pages 611-8

Zerbib D, Amalric F, Teissié J

Abstract

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.

MeSH Terms
Animals Cell Line Cell Separation Cell Transformation, Viral Cricetinae DNA Restriction Enzymes/metabolism DNA, Viral/analysis Deoxyribonuclease BamHI Deoxyribonuclease EcoRI Deoxyribonucleases, Type II Site-Specific Electric Stimulation Female Nucleic Acid Hybridization Plasmids Simplexvirus/genetics Thymidine Kinase/genetics Transfection
Chemicals
DNA, Viral Thymidine Kinase DNA Restriction Enzymes Deoxyribonuclease BamHI Deoxyribonuclease EcoRI CAGCTG-specific type II deoxyribonucleases Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Zerbib D
Amalric F
Teissié J
Article Info
Journal
Biochemical and biophysical research communications
Abbr.
Biochem Biophys Res Commun
ISSN
0006-291X
Published
1985-06-28
Pages
611-8
Language
English
Region
United States
NLM ID
0372516
Subset
IM
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