title: RNA decay by messenger RNA interferases.
abstract: Two abundant toxin-antitoxin (TA) gene families, relBE and mazEF, encode mRNA cleaving enzymes whose ectopic overexpression abruptly inhibits translation and thereby induces a bacteriostatic condition. Here we describe and discuss protocols for the overproduction, purification, and analysis of mRNA cleaving enzymes such as RelE of Escherichia coli and the corresponding antitoxin RelB. In particular, we describe a set of plasmid vectors useful for the detailed analysis of cleavage sites in model mRNAs.
authors.affiliation: Institute for Cell and Molecular Biosciences, Medical School, University of Newcastle, Newcastle, United Kingdom.
authors.forename: Mikkel
authors.initials: M
authors.lastname: Christensen-Dalsgaard
authors.forename: Martin
authors.initials: M
authors.lastname: Overgaard
authors.forename: Kristoffer Skovbo
authors.initials: KS
authors.lastname: Winther
authors.forename: Kenn
authors.initials: K
authors.lastname: Gerdes
mesh_terms: D001427:Bacterial Toxins / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D002852:Chromatography, Ion Exchange; D003062:Codon; D017931:DNA Primers; D029968:Escherichia coli Proteins / Q000235:genetics; D010957:Plasmids; D034622:RNA Interference; D015347:RNA Probes; D012329:RNA, Bacterial / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D001427:Bacterial Toxins; D003062:Codon; D017931:DNA Primers; D029968:Escherichia coli Proteins; D015347:RNA Probes; D012329:RNA, Bacterial; C465566:RelE protein, E coli