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. 1983 Feb;80(3):653–657. doi: 10.1073/pnas.80.3.653

Decay of mRNA in Escherichia coli: investigation of the fate of specific segments of transcripts.

A von Gabain, J G Belasco, J L Schottel, A C Chang, S N Cohen
PMCID: PMC393437  PMID: 6187001

Abstract

An assay was developed to investigate the fate of specific segments of beta-lactamase (bla) and ompA gene transcripts in Escherichia coli. DNA probes cloned in bacteriophage M13 were treated with an endonuclease capable of cleaving single-stranded DNA, the fragments produced were annealed with total cellular RNA, and the resulting RNA . DNA hybrids were subjected to S1 nuclease treatment and gel fractionation. By using this assay, direct evidence was obtained for 3'-to-5' directionality in the decay of the long-lived mRNA encoded by the ompA gene, and no preferential stability was observed for translated versus untranslated mRNA segments. In the case of bla mRNA, initial cleavage of the full-length transcript was rate limiting, and no decay intermediates were detected. No difference in degradation rate was seen for bla transcripts having variant 3' or 5' termini.

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