[支气管败血波氏杆菌细胞中百日咳毒素操纵子在其自身启动子和异源启动子控制下的表达特征]

[Features of expression of the pertussis toxin operon under the control of its own and heterologous promotors in Bordetella bronchiseptica cells].

作者信息

Govorun V M, Ivanova V F, Kirillov M Iu, Klitsunova N V, Nechaeva E V, Siniashina L N, Shumakov Iu L, Karataev G I

出版信息

Mol Gen Mikrobiol Virusol. 1993 Mar-Apr(2):21-7.

Abstract

Expression of the cloned operon encoding the pertussis toxin synthesis under the control of operons own vir-dependent promoter or vir-independent promoter of Escherichia coli origin was studied. Proteins produced by the recombinant strains have been characterized. The pertussis toxin operon was shown to express under the control of both homologous and heterologous promoters in Bordetella bronchiseptica cells. Use of the lac-promoter increases the yield of produced toxin twofold. Copy number of operon in the cell does not influence the level of toxin production. The synthesized protein can be transported into the culture medium. The biological and physico-chemical properties of the protein are similar to the ones of the natural pertussis toxin. Bordetella bronchiseptica strain producing the toxin with decreased toxic activity has been obtained. Thus, a simple system for cellular expression of the toxin operon was constructed in Bordetella bronchiseptica. It permits one to construct new strains producing nontoxic derivatives of the pertussis toxin for construction of nonreactogenic vaccines.

摘要

研究了在源自大肠杆菌的操纵子自身的毒力依赖性启动子或毒力非依赖性启动子控制下，编码百日咳毒素合成的克隆操纵子的表达。对重组菌株产生的蛋白质进行了表征。结果表明，百日咳毒素操纵子在支气管败血波氏杆菌细胞中在同源和异源启动子的控制下均能表达。使用乳糖启动子可使产生的毒素产量提高两倍。细胞中操纵子的拷贝数不影响毒素产生水平。合成的蛋白质可转运到培养基中。该蛋白质的生物学和物理化学性质与天然百日咳毒素相似。已获得产生毒性活性降低的毒素的支气管败血波氏杆菌菌株。因此，在支气管败血波氏杆菌中构建了一个用于毒素操纵子细胞表达的简单系统。它允许构建产生百日咳毒素无毒衍生物的新菌株，用于构建无反应原性疫苗。