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苏云金芽孢杆菌以色列变种晶体δ-内毒素:对昆虫和哺乳动物细胞的体外及体内作用

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

作者信息

Thomas W E, Ellar D J

出版信息

J Cell Sci. 1983 Mar;60:181-97. doi: 10.1242/jcs.60.1.181.

Abstract

Bacillus thuringiensis var israelensis parasporal crystal delta-endotoxin was purified by ultracentrifugation on a discontinuous sucrose gradient. Native delta-endotoxin crystals showed no detectable toxicity in the vitro and in vivo systems that are described. By contrast alkali-solubilized crystal delta-endotoxin caused rapid cytological and cytopathological changes in Aedes albopictus, Choristoneura 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 delta-endotoxin. In addition the soluble crystal delta-endotoxin protein caused haemolysis of rat, mouse, sheep, horse and human erythrocytes. Intravenous administration of the alkali-soluble crystal delta-endotoxin to Balb. c mice at a dose rate of 15-30 micrograms of protein per gram body weight resulted in rapid paralysis followed by death within 12h. Subcutaneous inoculation of 15-30 micrograms of protein per gram body weight resulted in death of suckling mice in 2-3 h. The alkali-solubilized crystal delta-endotoxin was not toxic however, when administered per os. A comparison is made with a similar alkali-soluble fraction from the parasporal crystal delta-endotoxin of B. thuringiensis var kurstaki. With the exception of the Lepidopteran cell line, Choristoneura fumiferana 63 CF1, this soluble crystal delta-endotoxin protein showed no in vitro or in vivo toxicity, and no haemolytic activity.

摘要

苏云金芽孢杆菌以色列变种的伴孢晶体δ-内毒素通过在不连续蔗糖梯度上进行超速离心来纯化。天然δ-内毒素晶体在所述的体外和体内系统中未显示出可检测到的毒性。相比之下,碱溶性晶体δ-内毒素在白纹伊蚊、烟色毒蛾63 CF1、草地贪夜蛾和粉纹夜蛾细胞系中引起了快速的细胞学和细胞病理学变化,这通过相差显微镜和活体染色观察到。小鼠成纤维细胞、原代猪淋巴细胞和三种小鼠上皮癌细胞类型对碱溶性晶体δ-内毒素表现出类似的反应。此外,可溶性晶体δ-内毒素蛋白导致大鼠、小鼠、绵羊、马和人红细胞溶血。以每克体重15 - 30微克蛋白质的剂量率将碱溶性晶体δ-内毒素静脉注射给Balb. c小鼠,导致快速麻痹,随后在12小时内死亡。每克体重皮下接种15 - 30微克蛋白质导致乳鼠在2 - 3小时内死亡。然而,碱溶性晶体δ-内毒素经口给药时无毒。将其与来自苏云金芽孢杆菌库尔斯塔克变种伴孢晶体δ-内毒素的类似碱溶性部分进行了比较。除了鳞翅目细胞系烟色毒蛾63 CF1外,这种可溶性晶体δ-内毒素蛋白在体外或体内均未显示出毒性,也没有溶血活性。

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