Figure 5. Effect of YoeBsl on protein synthesis in a cell-free system and Primer extension analysis of YoeBsl cleavage sites in the ompA mRNA in vivo.

A) MazG protein synthesis was carried out using E. coli T7 S30 extract system for circular DNA (Promega) with pET11a-mazG. Lane 1, without YoeB-His₆sl; lanes 2 to 5, 0.1, 0.2, 0.5 and 1 µM YoeB-His₆sl were added, respectively; lane 6, 1 µM YoeB-His₆sl plus 1 µM YefM-His₆sl; and lane 7, 1 µM YefM-His₆sl was added. B) Total RNA was prepared from E. coli BL21 cells harboring pFUS2-Tox at indicated time points before and after the induction of yoeBsl expression. The major cleavage site is indicated by an arrowhead on the right side of the gel. The sequence of the major cleavage site in the ompA mRNA is also shown on the right side of the gel. The sequence ladder for ompA was obtained using pCR®2.1-TOPO-ompA as template. The full-length RNA bands (FL) are indicated with an arrow.