RNA decay by messenger RNA interferases.
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.
Christensen-Dalsgaard
Mikkel
M
Institute for Cell and Molecular Biosciences, Medical School, University of Newcastle, Newcastle, United Kingdom.
Overgaard
Martin
M
Winther
Kristoffer Skovbo
KS
Gerdes
Kenn
K
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*
D001427:Bacterial Toxins; D003062:Codon; D017931:DNA Primers; D029968:Escherichia coli Proteins; D015347:RNA Probes; D012329:RNA, Bacterial; C465566:RelE protein, E coli