FIG. 6.
Northern blot analysis of the in vitro cleavage of sarA and recA mRNAs by MazFSa. The sarA and recA mRNAs were transcribed from BamHI-linearized pET14b-sarA and pET14b-recA plasmids by using a T7 large-scale transcription kit (Promega) and digested with purified MazFSa as described in Materials and Methods. Amounts of 5 μg of sarA and recA mRNAs were digested with 15 pmol MazFSa at times indicated for panels A and B. (A) sarA mRNA cleavage by MazFSa. Lane 1, no MazFSa; lanes 2 to 4, digestion with MazFSa for 60, 90, and 120 min, respectively. (B) recA mRNA cleavage by MazFSa. Lane 1, no MazFSa; lane 2, digestion with MazFSa for 60 min. The cleaved fragments were detected with the 32P-radiolabeled sarA and recA DNA probes in Northern blots as described in Materials and Methods.