title: Instability of extrachromosomal cosmid DNA in SV40-transformed human (ataxia-telangiectasia) cells.
abstract: The ability of SV40-transformed human (ataxia-telangiectasia) fibroblasts to maintain Epstein-Barr virus (EBV)-based plasmids and cosmids extrachromosomally has been investigated. Transfection of a culture of cells with two different plasmids gave rise to cell clones which were able to maintain both plasmids extrachromosomally. When an EBV-based cosmid library was transfected into the cells and an individual cell clone was isolated, the extrachromosomal DNA derived from the cosmid contained numerous deletions and rearrangements. When individual cosmids were transfected into the culture, and several cell clones were isolated, the intracellular cosmid-derived DNA again showed the presence of multiple deletions and rearrangements. We conclude that although SV40-transformed cells are able to maintain more than one different EBV-based plasmid extrachromosomally, large EBV-derived molecules are extensively rearranged. SV40-transformed human fibroblasts cannot therefore be usefully used in attempting to clone genes from EBV-based cosmid libraries.
authors.affiliation: MRC Cell Mutation Unit, University of Sussex, Brighton, United Kingdom.
authors.forename: S W
authors.initials: SW
authors.lastname: Dean
authors.forename: L
authors.initials: L
authors.lastname: Kincla
authors.forename: H R
authors.initials: HR
authors.lastname: Sykes
authors.forename: A R
authors.initials: AR
authors.lastname: Lehmann
authors.forename: I A
authors.initials: IA
authors.lastname: Wise
mesh_terms: D001260:Ataxia Telangiectasia; D002460:Cell Line; D002472:Cell Transformation, Viral; D003360:Cosmids; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D005814:Genes, Viral; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D010957:Plasmids; D013539:Simian virus 40 / Q000235:genetics*; D014162:Transfection
chemical_list: D004279:DNA, Viral; D004247:DNA