FIG. 4.

Western blot analysis of overexpressed Ncr-His₆ protein in E. coli and total cellular protein fractions of P. syringae. Pseudomonas cells were grown to stationary phase (OD₆₀₀ = 5.0) at 18, 22, and 28°C and then subjected to total protein extraction. Western blotting was performed with polyclonal antibodies raised against Ncr. The specific signal is marked with an arrow. Similar signals were detected in protein samples from cultures grown at 22°C (data not shown). Lanes: 1, crude extract of E. coli M15 (pREP4/pQE-Ncr); 2, crude extract of E. coli M15 (pREP4/pQE-Ncr) after induction with 0.1 mM IPTG; 3, purified Ncr-His₆ protein; 4, P. syringae pv. glycinea PG4180.N9 grown at 18°C; 5, PG4180.N9 grown at 28°C; 6, P. syringae pv. glycinea race 0 grown at 18°C; 7, P. syringae pv. glycinea race 5 grown at 18°C; M, molecular size marker.