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PMID: 8289382 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The genomic instability associated with integrated simian virus 40 DNA is dependent on the origin of replication and early control region.

Journal of virology ·Vol. 68 ·No. 2 ·1994-02-00 ·Pages 787-96

Hunter DJ, Gurney EG

Abstract

DNA rearrangements in the form of deletions and duplications are found within and near integrated simian virus 40 (SV40) DNA in nonpermissive cell lines. We have found that rearrangements also occur frequently with integrated pSV2neo plasmid DNA. pSV2neo contains the entire SV40 control region, including the origin of replication, both promoters, and the enhancer sequences. Linearized plasmid DNA was electroporated into X1, an SV40-transformed mouse cell line that expresses SV40 large T antigen (T Ag) and shows very frequent rearrangements at the SV40 locus, and into LMtk-, a spontaneously transformed mouse cell line that contains no SV40 DNA. Stability was analyzed by subcloning G-418-resistant clones and examining specific DNA fragments for alterations in size. Five independent X1 clones containing pSV2neo DNA were unstable at both the neo locus and the T Ag locus. By contrast, four X1 clones containing mutants of pSV2neo with small deletions in the SV40 core origin and three X1 clones containing a different neo plasmid lacking SV40 sequences were stable at the neo locus, although they were still unstable at the T Ag locus. Surprisingly, five independent LMtk- clones containing pSV2neo DNA were unstable at the neo locus. LMtk- clones containing origin deletion mutants were more stable but were not as stable as the X1 clones containing the same plasmid DNA. We conclude that the SV40 origin of replication and early control region are sufficient viral components for the genomic instability at sites of SV40 integration and that SV40 T Ag is not required.

MeSH Terms
Animals Antigens, Viral, Tumor/genetics Cloning, Molecular Electroporation Gene Rearrangement Genes, Reporter Genome, Viral Plasmids/genetics Regulatory Sequences, Nucleic Acid/genetics Sequence Deletion Simian virus 40/genetics,growth & development Transformation, Genetic Virus Integration/genetics Virus Replication/genetics
Chemicals
Antigens, Viral, Tumor
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Hunter D J
Department of Biology, University of Utah, Salt Lake City 84112.
Gurney E G
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.68.2.787-796.1994
Published
1994-02-00
Pages
787-96
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC236515
Subset
IM
Grants
NCI NIH HHS · 5 P30 CA42014 · United States
External Links
PubMed source DOI: 10.1128/JVI.68.2.787-796.1994 PMC Full Text View journal details
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