Figure 3

Increase of the messenger RNA inactivation rate with the MazF interferase in a cell-free system. (A) Addition of pure MazE into a MazF cell-free reaction (MazF extract). End-point measurements of Luc production (5 nM pBEST-UTR1-Luc) and deGFP production (5 nM pBEST-UTR1-eGFP). (B) End-point measurements of Luc synthesized in a standard cell-free reaction (no MazF) as the function of MazF added to the reaction (5 nM pBEST-UTR1-Luc). The last bar is a control experiment with 140 nM MazF and 1 μM MazE. (C) Kinetics of deGFP expression (1 nM pBEST-OR2-OR1-Pr-UTR1-deGFP-T500) with a range of MazF. The MazE sample was tested with 90 nM MazF. (D) Kinetics of deGFP expression (5 nM plasmid pBEST-OR2-OR1-Pr-UTR1-deGFP-T500) with 36 nM of MazF after addition of 30 μM Tagetin.