FIG. 5.
Localization of GerD in germinated spores. Spores of strain PP19 (gerD amyE::PsspB-gerD-flag) were germinated in LB medium plus 10 mM l-alanine for 45 min, and germinated spores were purified as described in Materials and Methods. The germinated spores were lyophilized, disrupted, extracted, and fractionated as described in Materials and Methods. Aliquots of fractions from equal amounts of spores were run by SDS-PAGE, proteins were transferred to a PVDF membrane, and GerD-FLAG was detected with anti-FLAG antibody. Samples run in the various lanes were as follows: 1, total extract; 2, insoluble fraction; 3, soluble fraction; 4, S100; and 5, P100. The numbered lines to the left of the figure denote the migration positions of molecular mass markers, in kDa, and the arrowheads to the right indicate the predicted migration positions of GerD-FLAG monomers (M), dimers (D), and trimers (T).