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PMID: 6264155 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Polyoma virus minichromosomes: a soluble in vitro replication system.

Journal of virology ·Vol. 38 ·No. 3 ·1981-06-00 ·Pages 805-14

Gourlie BB, Krauss MR, Buckler-White AJ, Benbow RM, Pigiet V

Abstract

Polyoma virus minichromosomes were isolated from infected 3T6 cells by hypotonic extraction of isolated nuclei. The kinetics of in vitro DNA synthesis in the nuclear extract was similar to that observed with intact nuclei. The majority of the products of in vitro DNA synthesis sedimented with replicative intermediate (RI) minichromosomes and migrated as two bands (RI-a and RI-b) on 1.4% agarose gels. The kinetics of deoxynucleotide monophosphate incorporation into these species was consistent with the existence of several rate-limiting steps in in vitro replication by polyoma minichromosomes. Electron microscope analysis showed that the RI-a band consisted almost entirely of RI theta structures ranging from 46 to 87% replicated, with one-half of all theta structures 67 +/- 4% replicated. The RI-b material was more complex, consisting of sigma and alpha structures with tails ranging from 7 to 114% of polyoma genome length and, less frequently, of linked and multiple linked dimeric structures.

MeSH Terms
Bacterial Proteins Centrifugation, Density Gradient DNA Restriction Enzymes DNA, Viral/biosynthesis Deoxyribonucleases, Type II Site-Specific Electrophoresis, Agar Gel Genes, Viral Microscopy, Electron Nucleic Acid Conformation Polyomavirus/genetics,metabolism Replicon
Chemicals
Bacterial Proteins DNA, Viral DNA Restriction Enzymes BglII endonuclease Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Gourlie B B
Krauss M R
Buckler-White A J
Benbow R M
Pigiet V
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30 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.38.3.805-814.1981
Published
1981-06-00
Pages
805-14
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC171217
Subset
IM
Grants
NCI NIH HHS · CA-06304 · United States
NIGMS NIH HHS · GM-23813 · United States
NIGMS NIH HHS · GM-23970 · United States
External Links
PubMed source DOI: 10.1128/JVI.38.3.805-814.1981 PMC Full Text View journal details
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