Fig 8.

Analysis of the level of biotinylation of GerKC in decoated dormant and germinated spores of C. perfringens wild-type strain SM101. Biotinylation was carried out on decoated dormant spores and intact germinated spores as described in Materials and Methods. Spores were disrupted and fractionated, and the levels of various germination proteins in (i) the total IM fraction (T), (ii) the IM fraction that did not adsorb to the NeutrAvidin beads (F), and (iii) the eluate from the NeutrAvidin beads (E) were assayed by Western blotting using anti-GerKC antiserum, as described in Materials and Methods. Aliquots in the T, F, and E samples were from equal amounts of spores. The F_(50%) sample (lane 4) from dormant spores was also the protein that did not adsorb to the NeutrAvidin beads but was from 1/2 the amount of spores as in the other lanes. The E_(50%) sample (lane 8) from germinated spores was also the eluate from the NeutrAvidin beads but was from 1/2 the amount of spores as in the other lanes. Analysis of this Western blot by the program ImageJ indicated that ∼33% of GerKC was biotinylated in dormant spores, with ∼50% biotinylated in germinated spores.