Detection of high levels of polyadenylate-containing RNA in bacteria by the use of a single-step RNA isolation procedure.
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.
D001407:Bacillus / Q000032:analysis*; D001412:Bacillus subtilis / Q000032:analysis; D002846:Chromatography, Affinity / Q000379:methods; D004926:Escherichia coli / Q000032:analysis*; D009693:Nucleic Acid Hybridization; D011061:Poly A / Q000302:isolation & purification*; D011072:Poly U; D012313:RNA / Q000302:isolation & purification*; D012329:RNA, Bacterial / Q000302:isolation & purification*; D012333:RNA, Messenger; D013171:Spores, Bacterial / Q000032:analysis; D014316:Tritium
D012329:RNA, Bacterial; D012333:RNA, Messenger; D014316:Tritium; D011061:Poly A; D011072:Poly U; D012313:RNA