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Article: Replication of pSV2-gpt in COS-1 cells: Stability of plasmid DNA in the presence and absence of biochemical selection

TitleReplication of pSV2-gpt in COS-1 cells: Stability of plasmid DNA in the presence and absence of biochemical selection
Authors
Issue Date1985
PublisherSpringer New York LLC.
Citation
Somatic Cell and Molecular Genetics, 1985, v. 11 n. 2, p. 167-176 How to Cite?
AbstractWe have previously demonstrated that COS-1 cell lines transformed by pSV2-gpt and maintained under biochemical selection replicate multiple copies of extrachromosomal plasmid DNA (1). We have now examined the replication and stability of this DNA in a representative cell line. In situ hybridization analyses revealed that intense replication of pSV2-gpt occurs in only a small subpopulation of cells and results from bursts of plasmid replication that occur periodically and spontaneously in the cell population. This suggests that COS-1 cells are only semipermissive for pSV2-gpt replication. No correlation was observed between levels of pSV2-gpt replication and the presence or absence of biochemical selection for the Gpt marker. However, growth of cells under nonselective conditions led to a rapid and progressive loss of pSV2-gpt DNA. This loss correlated with segregation of Gpt- revertants that lacked detectable plasmid sequences. Hence, maintenance of pSV2-gpt in the cell line was dependent on continuous biochemical selection. Stable replication of pSV2-gpt could be observed as late as four months after transfection, suggesting that this system might be useful for propagation of cloned DNA in COS-1 cells for extended periods of time. However, by nine months, extensive rearrangements of pSV2-gpt sequences were detected, indicating ultimate instability of the plasmid in the host cells.
Persistent Identifierhttp://hdl.handle.net/10722/44208
ISSN
2000 Impact Factor: 0.884
2005 SCImago Journal Rankings: 1.061
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorTsui, L-Cen_HK
dc.contributor.authorBreitman, MLen_HK
dc.date.accessioned2007-09-12T03:49:01Z-
dc.date.available2007-09-12T03:49:01Z-
dc.date.issued1985en_HK
dc.identifier.citationSomatic Cell and Molecular Genetics, 1985, v. 11 n. 2, p. 167-176en_HK
dc.identifier.issn0740-7750en_HK
dc.identifier.urihttp://hdl.handle.net/10722/44208-
dc.description.abstractWe have previously demonstrated that COS-1 cell lines transformed by pSV2-gpt and maintained under biochemical selection replicate multiple copies of extrachromosomal plasmid DNA (1). We have now examined the replication and stability of this DNA in a representative cell line. In situ hybridization analyses revealed that intense replication of pSV2-gpt occurs in only a small subpopulation of cells and results from bursts of plasmid replication that occur periodically and spontaneously in the cell population. This suggests that COS-1 cells are only semipermissive for pSV2-gpt replication. No correlation was observed between levels of pSV2-gpt replication and the presence or absence of biochemical selection for the Gpt marker. However, growth of cells under nonselective conditions led to a rapid and progressive loss of pSV2-gpt DNA. This loss correlated with segregation of Gpt- revertants that lacked detectable plasmid sequences. Hence, maintenance of pSV2-gpt in the cell line was dependent on continuous biochemical selection. Stable replication of pSV2-gpt could be observed as late as four months after transfection, suggesting that this system might be useful for propagation of cloned DNA in COS-1 cells for extended periods of time. However, by nine months, extensive rearrangements of pSV2-gpt sequences were detected, indicating ultimate instability of the plasmid in the host cells.en_HK
dc.languageengen_HK
dc.publisherSpringer New York LLC.en_HK
dc.relation.ispartofSomatic Cell and Molecular Genetics-
dc.rightsThe original publication is available at www.springerlink.comen_HK
dc.subject.meshCulture mediaen_HK
dc.subject.meshDna replicationen_HK
dc.subject.meshDna, recombinanten_HK
dc.subject.meshDna, viral - geneticsen_HK
dc.subject.meshPlasmidsen_HK
dc.titleReplication of pSV2-gpt in COS-1 cells: Stability of plasmid DNA in the presence and absence of biochemical selectionen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0740-7750&volume=11&issue=2&spage=167&epage=176&date=1985&atitle=Replication+of+pSV2-gpt+in+COS-1+cells:+Stability+of+plasmid+DNA+in+the+presence+and+absence+of+biochemical+selectionen_HK
dc.description.naturelink_to_subscribed_fulltexten_HK
dc.identifier.doi10.1007/BF01534705-
dc.identifier.pmid2984789-
dc.identifier.isiWOS:A1985AEF2000007-

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