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KMID : 1041219910330010032
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Journal of Sericultural and Entomological Science
1991 Volume.33 No. 1 p.32 ~ p.36
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Purtification of Parasporal Protein Crystals of Bacillus thuringiensis
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Abstract
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The study has been carried out to acquire some basic informations about Bacillus thuringiensis for developing the microbial pesticide. Three strains of Bacillus thuringiensis var. kurstaki, dendrolimus and aizawai, were used in the experiments as follows. Growth characteristics of each strain were examined and parasporal protein crystals were isolated from the mixtures of spores and protein crystals by the new method of centrifugation in two-layer cushion of Renograffin using a fixed angle rotor. The results are as follows. 1. No difference was shown in growth characterestics among three strains of B. thuringiensis. In growth curves, all strains reached to exponential phase by 3 hr and stationary phase by 7-8 hr after inoculation. 2. The pH of the culture media during exponential growth stage decreased about 1.4 of a pH unit at the beginning of sporulation, but recovered during the early stage of sporulation and then remained nearly constant during the later stage. 3. As 10 ml sample was applied to two-layer cushion of Renograffin and then centrifuged for 1 hr at 27,000g a fixed angle rotor, the purity and recovery ratio was 99.9% and 5.8%, respectively. It has been shown that the new method for the isolation of parasporal protein crystals was more efficient than any from the estabilished methods.
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