Effect of YoeB on cellular mRNAs and identification of YoeB cleavage
sites in the chromosomally encoded lpp and ompA mRNAs.
Total cellular RNA was extracted from E. coli BW25113 cells
containing pBAD-YoeB at various time points as indicated after the addition of
arabinose and subjected to Northern blot analysis using radiolabeled
lpp and ompA open reading frames as probes shown in
A and B, respectively. For in vivo primer
extension, the lpp and ompA mRNAs were prepared from E.
coli BW25113 cells containing pBAD-YoeB at various time points as
indicated in C and D before and after the induction of YoeB.
The sequence ladders for lpp and ompA were obtained using
pCR®2.1-TOPO®-lpp and pCR®2.1-TOPO-ompA as
template, respectively. The sequences around the major cleavage sites are
shown at the right-hand side and the major primer extension stop
sites are indicated by arrowheads. On the top of the gels,
the full-length (FL) RNA bands are shown with an arrowhead
and FL. C, primer extension analysis of the lpp
mRNA. D, primer extension analysis of the ompA mRNA.
FL, full-length.