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PMID: 6169008 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Detection of high levels of polyadenylate-containing RNA in bacteria by the use of a single-step RNA isolation procedure.

Nucleic acids research ·Vol. 9 ·No. 14 ·1981-07-24 ·Pages 3545-54

Gopalakrishna Y, Langley D, Sarkar N

Abstract

A new one-step procedure for the isolation of bacterial RNA, involving lysis by proteinase K in the presence of sodium dodecyl sulfate, is described. Pulse-labeled RNA isolated by this procedure for Bacillus brevis, Bacillus subtilis, and Escherichia coli B has been found to contain a substantial fraction (15-40%) of polyadenylated RNA as determined by adsorption to oligo(dT)-cellulose. This contrasts with RNA isolated by procedures involving phenol extraction, a process which appears to lead to the selective loss of polyadenylated RNA. The presence of polyadenylated RNA in E. coli was confirmed by an independent method which involved hybridization with [3H]polyuridylic acid. Using the proteinase K method for RNA isolation, it was possible to demonstrate the in vitro synthesis of polyadenylated RNA by toluene-treated cells of B. brevis, B. subtilis, and E. coli.

MeSH Terms
Bacillus/analysis Bacillus subtilis/analysis Chromatography, Affinity/methods Escherichia coli/analysis Nucleic Acid Hybridization Poly A/isolation & purification Poly U RNA/isolation & purification RNA, Bacterial/isolation & purification RNA, Messenger Spores, Bacterial/analysis Tritium
Chemicals
RNA, Bacterial RNA, Messenger Tritium Poly A Poly U RNA
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Gopalakrishna Y
Langley D
Sarkar N
References (24)
24 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/9.14.3545
Published
1981-07-24
Pages
3545-54
Language
English
Region
England
NLM ID
0411011
PMCID
PMC327370
Subset
IM
Grants
NIGMS NIH HHS · GM 26517 · United States
External Links
PubMed source DOI: 10.1093/nar/9.14.3545 PMC Full Text View journal details
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