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PMID: 1508725 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

RecA protein promotes rapid RNA-DNA hybridization in heterogeneous RNA mixtures.

Nucleic acids research ·Vol. 20 ·No. 16 ·1992-08-25 ·Pages 4347-53

Kirkpatrick DP, Radding CM

Abstract

The nucleoprotein filament formed by the RecA protein of Escherichia coli on single-stranded DNA catalyzes the hybridization of RNA transcripts with single-stranded DNA sequences at 37 degrees C, in vitro. RecA protein rapidly promotes hybridization, even when noncomplementary RNA is in a millionfold nucleotide excess over hybridizing RNA, and in a thousandfold nucleotide excess over hybridizing single-stranded DNA. Heterologous double-stranded DNA and RecA-coated noncomplementary single-stranded DNA are also poor competitors of RNA transcripts produced in vitro. Since large excesses of noncomplementary RNA fail to inhibit sharply the hybridization reaction by RecA protein under mild, non-degradative conditions, the reaction may be useful in the identification and isolation of transcripts produced in vivo.

MeSH Terms
DNA, Single-Stranded/metabolism Electrophoresis HeLa Cells Humans Nucleic Acid Hybridization/physiology RNA, Messenger/metabolism RNA, Ribosomal, 16S/metabolism RNA, Ribosomal, 23S/metabolism Rec A Recombinases/metabolism
Chemicals
DNA, Single-Stranded RNA, Messenger RNA, Ribosomal, 16S RNA, Ribosomal, 23S Rec A Recombinases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kirkpatrick D P
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT 06510.
Radding C M
References (12)
12 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1992-08-25
Pages
4347-53
Language
English
Region
England
NLM ID
0411011
PMCID
PMC334146
Subset
IM
Grants
NHGRI NIH HHS · HG00338-02 · United States
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