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. Author manuscript; available in PMC: 2014 Dec 1.
Published in final edited form as: Anal Biochem. 2013 Aug 29;443(1):10.1016/j.ab.2013.08.018. doi: 10.1016/j.ab.2013.08.018

Fig. 2.

Fig. 2

Cleavage of an mRNA transcript substrate by MazFSa(His)6. A transcript was produced as described in Materials and Methods. Incubation of the full length transcript (1070 nucleotides [nts]) with MazFSa(His)6 resulted in the cleavage products shown in the gel and depicted in the schematic below the gel. Left: Cleavage of the 3-site substrate (50 nM) produced 4 cleavage products within 240 s incubation with MazFSa(His)6 (50 nM). The products from MazFSa(His)6 cleavage at site 1 (UACAU) and site 3 (UACGU) are depicted below the gel. MazFSa(His)6 did not cleave at site 2 (UACGU) within 240 s. Right: Cleavage of the 1-site substrate under the same conditions results in two well-separated products. As shown below the gel, in the 1-site substrate the original cleavage sites 1 and 2 were removed and site 3 was changed to the optimal MazFSa recognition sequence, UACAU. Products were separated by denaturing gel electrophoresis and visualized by ethidium bromide staining.