Cloning and sequence analysis of a newly identified Brucella abortus gene and serological evaluation of the 17-kilodalton antigen that it encodes
Cloning and sequence analysis of a newly identified Brucella abortus gene and serological evaluation of the 17-kilodalton antigen that it encodes
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新鉴定的流产布鲁氏菌基因的克隆和序列分析及其编码的 17 千道尔顿抗原的血清学评估
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发表时间：
1995
期刊：
影响因子：
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通讯作者：
Eric Saman
中科院分区：
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作者：
Fabienne Hemmen;V. Weynants;Thierry Scarcez;J. Letesson;Eric Saman
迄今为止，编码流产布鲁氏菌蛋白的未知基因已从用布鲁氏菌感染绵羊血清探测的 lambda gt11 表达文库中分离出来。克隆基因的序列分析显示存在一个由 158 个氨基酸组成的开放阅读框，编码 17.3 kDa（计算分子量）的蛋白质。在大肠杆菌中表达的重组流产布鲁氏菌蛋白和相应的羊种布鲁氏菌蛋白以相同的表观分子质量迁移，如蛋白质印迹（免疫印迹）所示。在来自羊种布鲁氏菌或流产布鲁氏菌感染的绵羊和牛的一系列血清样品中，在对布鲁氏菌总蛋白提取物进行蛋白质印迹分析时，分别有 51% 和 39% 显示出 17 kDa 的信号。在使用特定单克隆抗体的竞争性酶联免疫吸附测定中，绵羊和牛的这一数字分别为 70% 和 61%。这些数据表明17-kDa抗原可用于布鲁氏菌感染的血清学诊断。
A thus far unknown gene encoding a Brucella abortus protein has been isolated from a lambda gt11 expression library probed with sera from Brucella-infected sheep. Sequence analysis of the cloned gene revealed the presence of an open reading frame of 158 amino acids encoding a protein of 17.3 kDa (calculated molecular mass). The recombinant B. abortus protein, expressed in Escherichia coli, and the corresponding Brucella melitensis protein migrated at the same apparent molecular masses as shown by Western blotting (immunoblotting). Among a series of serum samples from B. melitensis- or B. abortus-infected sheep and cows, 51 and 39%, respectively, showed a signal at 17 kDa on Western blot analysis of total protein extract from Brucella bacteria. These figures amount to 70 and 61% for sheep and cattle, respectively, in a competitive enzyme-linked immunosorbent assay with a specific monoclonal antibody. These data indicate that the 17-kDa antigen may be useful for serological diagnosis of Brucella infection.