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PMID: 2995921 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A novel transcription property of SP6 and T7 RNA polymerases: dependence on template structure.

Nucleic acids research ·Vol. 13 ·No. 17 ·1985-09-11 ·Pages 6223-36

Schenborn ET, Mierendorf RC

Abstract

The in vitro synthesis of extraneous RNA sequences by SP6 and T7 RNA polymerases from specific DNA templates is described. Transcription of templates prepared by digestion with restriction enzymes that leave 3' protruding ends resulted in the production of significant amounts of long, template-sized RNA transcripts which hybridized to vector DNA. Sequences copied from the noncoding template strand were among the extraneous transcripts. The presence of these sequences in probe preparations were detected in Southern and RNase protection hybridization assays. In contrast, transcription of DNA templates with blunt or 5' protruding ends yielded few RNA products as extraneous sequences.

MeSH Terms
Base Sequence Coliphages/enzymology,genetics DNA Restriction Enzymes DNA, Recombinant/metabolism DNA-Directed RNA Polymerases/metabolism Escherichia coli/enzymology,genetics Kinetics Plasmids T-Phages/enzymology,genetics Templates, Genetic Transcription, Genetic
Chemicals
DNA, Recombinant DNA-Directed RNA Polymerases DNA Restriction Enzymes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Schenborn E T
Mierendorf R C
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21 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/13.17.6223
Published
1985-09-11
Pages
6223-36
Language
English
Region
England
NLM ID
0411011
PMCID
PMC321948
Subset
IM
Grants
NCI NIH HHS · PCM-8309618 · United States
External Links
PubMed source DOI: 10.1093/nar/13.17.6223 PMC Full Text View journal details
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