New host cell system for regulated simian virus 40 DNA replication

R. D. Gerard, Y. Gluzman

Research output: Contribution to journalArticle

91 Citations (Scopus)

Abstract

Transformed monkey cell lines (CMT and BMT) that inducibly express simian virus 40 (SV40) T antigen from the metallothionein promoter have been isolated and characterized. Immunoprecipitaition of pulse-labeled T antigen demonstrates a 5- to 12-fold increase in the rate of syntheses on addition of heavy-metal inducers to the culture medium. Radioimmunoassay of cell extracts indicates the accumulation of three- to four-fold more total T antigen after 2 days of induction by comparison with uninduced controls. A direct correlation was found between the level of T-antigen synthesis and the extent of SV40 DNA replication in inducible cells. Inducible BMT cells expressing a low basal level of T antigen were efficiently transformed by a vector carrying the neomycin resistance marker and an SV40 origin of replication. These vector sequences were maintained in an episomal form in most G418-resistant cell lines examined and persisted even in the absence of biochemical selection. Extensive rearrangements were observed only if the vector contained bacterial plasmid sequences. Expression of a protein product under the control of the SV40 late promoter in such vectors was increased after heavy-metal-dependent amplification of the template. These results demonstrate the ability of BMT cells to maintain a cloned eucaryotic gene in an amplifiable episomal state.

Original languageEnglish (US)
Pages (from-to)3231-3240
Number of pages10
JournalMolecular and Cellular Biology
Volume5
Issue number11
StatePublished - 1985

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Simian virus 40
Viral Tumor Antigens
DNA Replication
Heavy Metals
Transformed Cell Line
Replication Origin
Neomycin
Metallothionein
Cell Extracts
Radioimmunoassay
Haplorhini
Culture Media
Plasmids
Cell Line
Genes
Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

New host cell system for regulated simian virus 40 DNA replication. / Gerard, R. D.; Gluzman, Y.

In: Molecular and Cellular Biology, Vol. 5, No. 11, 1985, p. 3231-3240.

Research output: Contribution to journalArticle

Gerard, RD & Gluzman, Y 1985, 'New host cell system for regulated simian virus 40 DNA replication', Molecular and Cellular Biology, vol. 5, no. 11, pp. 3231-3240.
Gerard, R. D. ; Gluzman, Y. / New host cell system for regulated simian virus 40 DNA replication. In: Molecular and Cellular Biology. 1985 ; Vol. 5, No. 11. pp. 3231-3240.
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