Detection of high levels of polyadenylate-containing RNA in bacteria by the use of a single-step RNA isolation procedure.
AUTOR(ES)
Gopalakrishna, Y
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=327370Documentos Relacionados
- Turnover of polyadenylate-containing ribonucleic acid in Saccharomyces cerevisiae.
- Identification of Discrete Polyadenylate-containing RNA Components Transcribed from HeLa Cell Mitochondrial DNA*
- A simple single-step procedure for small-scale preparation of Escherichia coli plasmids.
- Rate of Synthesis of Polyadenylate-Containing Ribonucleic Acid During the Yeast Cell Cycle
- Confirmation of the single-step membrane filtration procedure for estimating Pseudomonas aeruginosa densities in water.