Fig 4.

Effect of overproducing a functional ParEC fusion protein on the growth of RFM475 and CT170 strains. (a) The effect of pET11-parEC on the growth of RFM475 and CT170 cells was monitored by spotting 10 μl of serial 10-fold dilutions of cells grown in LB to an OD₆₀₀ of 0.6 (at 30 and 37°C, respectively, for RFM475 and CT170) from 10⁰ to 10⁻⁵ (from left to right) on LB plates that were incubated at 30°C for 48 h (RFM475) or 37°C for 24 h (CT170). Cells grown in liquid were also used to calculate the efficiency of plating (EOP; number of viable cells [colonies] without plasmid divided by the number of viable cells carrying pET11-parEC) and the doubling time. The results shown here are representative of three independent experiments. (b) CT170 and RFM475 cells with or without pET11-parEC were grown on LB plates at 37°C for 24 h. Aliquots of cells were recovered for Western blotting with anti-ParC (top) or anti-ParE (bottom) antibodies as described in Materials and Methods.