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


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1985-03

AUTHORS

Lap-Chee Tsui, Martin L. Breitman

ABSTRACT

We 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. More... »

PAGES

167-176

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf01534705

DOI

http://dx.doi.org/10.1007/bf01534705

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1028697615

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/2984789


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