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[利用重组蛋白对突尼斯金蝎毒液致死活性进行体内和体外保护]

[In vivo and in vitro protection against lethal activity of Buthus occitanus tunetanus venom with a recombinant protein].

作者信息

Benkhadir K, Mejri T, Bel Haj Rhouma R, El Ayeb M, Karoui H

机构信息

Laboratoire des Venins et Toxines, Institut Pasteur de Tunis, Tunis-Belvedère.

出版信息

Arch Inst Pasteur Tunis. 2002;79(1-4):19-26.

Abstract

We report the use of recombinant scorpion toxin in the form of fusion protein as antigen for mice immunisation. The aim is to produce protective antisera against lethal activity of the venom from Tunisian scorpion Buthus occitanus tunetanus, responsible for several annually reported human cases of scorpion stings. The gene encoding Bot III (the most toxic alpha toxin of Buthus occitanus tunetanus) was fused to the sequence encoding synthetic ZZ domains of staphylococcal protein A. The construct ZZ-Bot III was expressed in the periplasm of E. coli as a fusion protein and purified by affinity chromatography. The recombinant fusion protein was characterized and used as antigen to generate antibodies in mice. The antibodies against the recombinant protein neutralize the toxic venom (10 LD50/ml) and also confer protection for immunized mice against antigenically related mammal toxins.

摘要

我们报道了使用融合蛋白形式的重组蝎毒素作为抗原对小鼠进行免疫。目的是制备针对突尼斯蝎Buthus occitanus tunetanus毒液致死活性的保护性抗血清,该毒液每年导致多起人类蝎蜇伤病例。编码Bot III(Buthus occitanus tunetanus毒性最强的α毒素)的基因与编码葡萄球菌蛋白A合成ZZ结构域的序列融合。构建体ZZ-Bot III在大肠杆菌周质中作为融合蛋白表达,并通过亲和层析纯化。对重组融合蛋白进行了表征,并用作抗原在小鼠中产生抗体。针对重组蛋白的抗体可中和有毒毒液(10 LD50/ml),还能为免疫小鼠提供针对抗原相关哺乳动物毒素的保护。

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