FIG. 3.

Western blots showing α subunits formed by JM109(DE3) carrying pDTG141 derivatives with mutations as indicated below. A monoclonal antibody specific for the α subunit of NDO was used as described in Materials and Methods. Lanes: M, prestained molecular mass markers (Bio-Rad Laboratories, Hercules, Calif.); 1, purified wild-type NDO (2 μg); 2, F202L; 3, F202V; 4, V260A; 5, V260L; 6, V260N; 7, N201Q; 8, N201A; 9, N201S; 10, W316A; 11, T351N; 12, T351R; 13, T351S; 14, F352L; 15, F352V; 16, W358A; 17, D362A; 18, M366W; 19, pT7-5 (negative control); 20, wild-type NDO (pDTG141).