Figure 5
Cross-species complementation - both PP4397 and E. coli YcgR restore a motility defect to the double PP2258/PP4397 null derivative of P. putida. (A) Relative swimming motility of E. coli MG1655-ΔyhjH/ΔycgR carrying araC/PBAD expression plasmid for P. putida PP4397-FLAG (pVI2370) or E. coli YcgR-FLAG (pVI2373), assayed on 0.3% soft agar LB motility plates containing the indicated amount of L-arabinose as inducer. Graphed values are averages with standard deviations calculated from three independent colonies and were normalized by setting the values of the double mutant harbouring the vector control as 1. P-values, calculated with two-tailed student t-test (***P < 0.001), are relative to the vector control. Images are of corresponding protein expression levels as revealed by separation of 10 µg of soluble protein and subsequent Western analysis. Cropped images are derived from the same experiment processed in parallel on the same gel, and are shown alongside molecular size markers in Fig. S4B. (B) Relative swimming motility of P. putida KT2701-Δpp2258/Δpp4397, carrying araC/PBAD expression plasmid for PP4397-FLAG (pVI2370) or YcgR-FLAG (pVI2373) as under panel A. Full images of the corresponding Western analysis for the cropped images are shown alongside molecular size markers in Fig. S4B.
