FIG 5.

Western blot analysis of levels of MalS-GFP protein during spore germination. Dormant AR71 spores were germinated, protein was extracted from spores harvested before (t = 0) and after (t = 5, 15 min) germinant addition, spores were harvested and extracted, samples were run by the use of gel electrophoresis, and Western blot analysis carried out, all as described in Materials and Methods. For day 2 (young) spores prepared on 2× SG medium, 15 μg of protein was loaded on gels; for day 4 (old) spores, 30 μg of protein was loaded; and for day 2 (young) and day 4 (old) spores prepared in MOPS medium, 30-μg volumes of protein representative of all time points were loaded on gels. Spores from liquid medium were phase bright at t = 0 and 5 min and completed germination at 15 min. Spores obtained from solid medium completed germination at t = 5 min.