Crystals of a local isolate of Bacillus thuringiensis variety israelensis were isolated by centrifugation on a continuous sucrose gradient (40–70%). Analysis of the crystals by SDS-PAGE revealed three major protein subunits of Mr˜25,000, ˜66,000 and Mr˜140,000. The crystals were solubilised using high pH and reducing conditions. Analysis of the soluble (protoxin) and insoluble fractions by SDS-PAGE showed the presence of proteins of Mr˜21,000 and ˜61,000, respectively. Proteolytic treatment of these fractions resulted in no apparent change in the molecular weights of the proteins. The crystals contained carbohydrate moieties as determined by periodic acid Schiff (PAS) and fluorescein isothiocyanate (FITC) Concanavalin A staining. In double radial immunodiffusion experiments, antisera raised against the Mr˜21,000 and ˜66,000 subunits did not react with other B. thuringiensis protoxins known to be active against the tsetse fly, Glossina morsitans morsitans, and the stemborer, Chile partellus.
