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PMID: 4372611 Published · ppublish English Journal Article

Viral DNA in transformed cells. III. The amounts of different regions of the SV40 genome present in a line of transformed mouse cells.

Botchan M, Ozanne B, Sugden B, Sharp PA, Sambrook J

Abstract

(32)P-Labeled SV40 DNA was treated sequentially with restricting endonucleases EcoRI and Hpa I, and the resulting four fragments of DNA were separated by gel electrophoresis. The kinetics of renaturation of each of the fragments and of complete SV40 DNA were measured in the presence of DNA extracted from the SVT2 line of SV40-transformed mouse cells. It was found that these cells contain about six copies of a segment of DNA which includes the early region of the SV40 genome, and about one copy of the late viral sequences. To map the region of the viral genome which is transcribed in SVT2 cells, separated strands of each of the four fragments were prepared and hybridized to total transformed cell RNA. Part of the E strands of the two DNA fragments (A and C) which span the early region of the SV40 genome were found to enter the hybrid.

MeSH Terms
Animals Base Sequence Cell Line Cell Transformation, Neoplastic Chromosome Mapping Clone Cells DNA, Viral/analysis Diploidy Endonucleases Fibroblasts Genes Hydrolysis Kinetics Mice Nucleic Acid Denaturation Nucleic Acid Hybridization Nucleic Acid Renaturation Phosphorus Radioisotopes Polynucleotides/analysis RNA, Viral/analysis Simian virus 40
Chemicals
DNA, Viral Phosphorus Radioisotopes Polynucleotides RNA, Viral Endonucleases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Botchan M
Ozanne B
Sugden B
Sharp P A
Sambrook J
References (14)
14 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1974-10-00
Pages
4183-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC434354
Subset
IM
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