Vodkin M H, Williams J C
Rickettsial Diseases Laboratory, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland 21701-5011.
J Bacteriol. 1988 Mar;170(3):1227-34. doi: 10.1128/jb.170.3.1227-1234.1988.
A gene library from the DNA of Coxiella burnetii has been constructed in the cosmid vector pHC79. A particular clone, pJB196, reacted strongly with Coxiella-specific antibodies elicited in a number of different species of animals. This clone produced two abundant C. burnetii-specific polypeptides, a 14-kilodalton nonimmunoreactive protein and a 62-kilodalton immunoreactive protein. Sequencing identified two open reading frames, encoding polypeptides of 10.5 and 58.3 kilodaltons. The only transcriptional control element observed on the 5' side of the initiation codon resembled a heat shock promoter. This heat shock promoter was functionally regulated in Escherichia coli, since both proteins were produced by growth conditions at 37 degrees C and neither protein was detected at 23 degrees C. There were four sequences from the literature that were highly homologous (greater than 50%) to the 62-kilodalton protein from C. burnetii. Three were from Mycobacterium species and represent the immunodominant antigen of this genus. The other was from E. coli, detected as a gene that complements or suppresses a temperature-sensitive RNase activity. Since the recombinant protein was immunogenic, it may serve as an efficacious vaccine against C. burnetii and other pathogenic microorganisms that express the conserved antigen.
已使用黏粒载体pHC79构建了来自伯氏考克斯氏体DNA的基因文库。一个特定的克隆pJB196与多种不同动物体内产生的考克斯氏体特异性抗体发生强烈反应。该克隆产生了两种丰富的伯氏考克斯氏体特异性多肽,一种14千道尔顿的非免疫反应性蛋白和一种62千道尔顿的免疫反应性蛋白。测序鉴定出两个开放阅读框,分别编码10.5千道尔顿和58.3千道尔顿的多肽。在起始密码子5'侧观察到的唯一转录控制元件类似于热休克启动子。这种热休克启动子在大肠杆菌中受到功能调控,因为这两种蛋白质都是在37℃的生长条件下产生的,而在23℃时均未检测到任何一种蛋白质。文献中有四个序列与伯氏考克斯氏体的62千道尔顿蛋白高度同源(大于50%)。其中三个来自分枝杆菌属物种,代表该属的免疫显性抗原。另一个来自大肠杆菌,被检测为一个互补或抑制温度敏感型核糖核酸酶活性的基因。由于重组蛋白具有免疫原性,它可能作为一种有效的疫苗,用于预防伯氏考克斯氏体以及其他表达保守抗原的致病微生物。