Comparisons of Cells, Refractile Bodies, and Spores of Bacillus popilliae1

Brij M. Mitruka, Ralph N. Costilow, S. H. Black, R. E. Pepper
1967 Journal of Bacteriology  
The culture was maintained in broth as described previously (4). Cells were produced in a 1.5%lo Trypticase (BBL), 0.570 yeast extract, 0.2%70 glucose, and 0.6% K2HP04 medium (TYG) at pH 7.4, as described previously (8). Refractile bodies were produced in the "sporulation medium" as described by Costilow et al. (8) ; this medium is now designated TB medium. Unless otherwise specified, the inoculated medium was supplemented with 0.02%-70 DL-sodium-fB-hydroxybutyrate after 3 days of incubation,
more » ... ys of incubation, since this was found to stimulate the formation of refractile bodies (Mitruka and Costilow, un"published data). The refractile bodies were separated from the residual cells by centrifuging at 3,000 X g for 15 min as described previously (8 ). The spores harvested from hemolymph from larvae were washed six times and the vegetative cells and refractile bodies were washed twice with distilled water. Cell extracts were prepared with the Nossal cell 759 on March 22, 2020 by guest http://jb.asm.org/ Downloaded from MITRUKA ET AL. disintegrator (McDonald Engineering Co., Cleveland, Ohio) as described previously (25). Analytical methods. Proteins were estimated on 1 N NaOH extracts (40 C for 2 hr) of hot (90 C, 30 min) 760
doi:10.1128/jb.94.3.759-765.1967 fatcat:47tvjiyhvrcuvcr3npr6pknkwi