FIG 7.
Western blot analysis of FlgM expression in L. oakridgensis using an anti-FlgM antiserum. (A) Western blot of cell extracts of L. oakridgensis (Loa WT) or its isogenic ΔfliA mutant (Loa ΔfliA) grown to the exponential (E), postexponential (PE), or stationary (S) phase of growth and of E. coli (DH5α pBCSK, vector control) and E. coli expressing FlgM of L. oakridgensis (DH5α pflgMLoa). (B) Western blot of whole-cell lysates of L. oakridgensis harboring pKH12 grown at 30°C or 37°C and of supernatant of L. oakridgensis harboring plasmid pKH12. Samples were harvested at the exponential (E), postexponential (PE), and stationary (S) phases of growth (Fig. 3B), and equal amounts of bacterial lysates were loaded onto the gel (see Materials and Methods). The supernatant was 25-fold concentrated, and 15 μl was loaded onto the gel.