Bacillus thuringiensis var israelensis crystal δ-endotoxin: effects on insect and mammalian cells in vitro and in vivo

Journal of Cell Science - Tập 60 Số 1 - Trang 181-197 - 1983
Wendy E. Thomas1, David J. Ellar1
1University of Cambridge, Department of Biochemistry, Tennis Court Road, Cambridge CB2 1QW U.K.

Tóm tắt

ABSTRACT Bacillus thuringiensis var israelensis parasporal crystal ô-endotoxin was purified by ultracentrifugation on a discontinuous sucrose gradient. Native δ-endotoxin crystals showed no detectable toxicity in the in vitro and in vivo systems that are described. By contrast alkali-solubilized crystal δ-endotoxin caused rapid cytological and cytopathological changes in Aedes albopictus, Choris-toneura fumiferana 63 CF1, Spodoptera frugiperda and Trichoplusia ni cell lines as observed by phase-contrast microscopy and vital staining. Mouse fibroblasts, primary pig lymphocytes and three mouse epithelial carcinoma cell types showed a similar response to the alkali-soluble crystal δ-endotoxin. In addition the soluble crystal δ -endotoxin protein caused haemolysis of rat, mouse, sheep, horse and human erythrocytes. Intravenous administration of the alkali-soluble crystal δ-endotoxin to Balb.c mice at a dose rate of 15-30μg of protein per gram body weight resulted in rapid paralysis followed by death within 12h. Subcutaneous inoculation of 15–30μg of protein per gram body weight resulted in death of suckling mice in 2-3 h. The alkali-solubilized crystal δ-endotoxin was not toxic however, when administered per os. A comparison is made with a similar alkali-soluble fraction from the parasporal crystal δ-en-dotoxin of B. thuringiensis var kurstaki. With the exception of the Lepidopteran cell line, Choris-toneura fumiferana 63 CF1, this soluble crystal δ-endotoxin protein showed no in vitro or in vivo toxicity, and no haemolytic activity.

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Journal of Cell Science

Tập 60 Số 1

181-197

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