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PMID: 2981338 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

A soluble transcription system derived from purified vaccinia virions.

Journal of virology ·Vol. 53 ·No. 1 ·1985-01-00 ·Pages 205-13

Golini F, Kates JR

Abstract

A soluble extract from purified vaccinia virus particles has been developed which displays site-specific initiation of transcription on exogenous DNA templates that carry cloned vaccinia virus early gene sequences. Bacterial plasmid vectors with segments of a strongly expressed early region of the vaccinia virus genome were active templates, whether in supercoiled or linear, truncated forms. Correct initiation, corresponding to that found in vivo, was observed for all early genes tested. The involvement of other factors besides the viral RNA polymerase was demonstrated by the loss of specific initiation upon partial purification of the enzyme. Initiation activity was restored by reconstitution of the system with factors lacking polymerase activity. The soluble system retained properties of transcription characteristic of intact viral cores, including (i) similar relative rates of initiation of various genes, (ii) multiple requirement for ATP, (iii) methylation and polyadenylation of transcripts, and (iv) inhibition by a topoisomerase antagonist.

MeSH Terms
Base Sequence Cloning, Molecular DNA Restriction Enzymes Endonucleases Genes, Viral HeLa Cells Humans Kinetics Single-Strand Specific DNA and RNA Endonucleases Templates, Genetic Transcription, Genetic Vaccinia virus/genetics Virion/genetics
Chemicals
Endonucleases DNA Restriction Enzymes Single-Strand Specific DNA and RNA Endonucleases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Golini F
Kates J R
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38 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.53.1.205-213.1985
Published
1985-01-00
Pages
205-13
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC255011
Subset
IM
Grants
NIAID NIH HHS · AI 15941 · United States
External Links
PubMed source DOI: 10.1128/JVI.53.1.205-213.1985 PMC Full Text View journal details
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