Use of dominant repressor alleles for non-antibiotic selection in Yersinia pestis

描述(由申请人提供)： 鼠疫耶尔森氏菌是腺鼠疫、肺炎鼠疫和败血症鼠疫的病原体，目前作为潜在的生物恐怖主义武器而备受关注。在美国，如果用雾化鼠疫杆菌攻击，就会导致致命的肺炎形式，这将给人类的生命和经济带来严重的后果。在非洲、亚洲和美洲也继续自然地发生零星的鼠疫暴发。最近分离出了对多种抗生素具有抗药性的鼠疫耶尔森氏菌菌株，这表明鼠疫的自然爆发可能会变得更加难以控制。目前，没有有效的疫苗来预防鼠疫，除非在感染后立即给予抗生素，否则这种疾病通常是致命的。确定控制鼠疫杆菌的新策略的研究将需要对病原体进行基因操作。然而，目前使用的大多数技术都需要引入携带抗生素耐药性标记的遗传元素。具有抗生素耐药性的鼠疫杆菌不仅对研究人员构成了健康问题，而且如果耐药菌株被生物恐怖分子获得，也会增加风险。这项研究试图验证一个主要假设，即突变的阻遏蛋白将通过赋予对强有力的代谢抑制剂的抗性而作为可选择的标记发挥作用。这些研究将为鼠疫杆菌的基因操作提供新的工具，从而在不增加毒力的情况下更好地了解其致病性以及生产有效的疫苗。这里描述的方法也将适用于其他选定的生物恐怖主义制剂。

DESCRIPTION (provided by applicant): Yersinia pestis is the causative agent of bubonic, pneumonic and septicemic plague, and is currently a major concern as a potential bioterrorism weapon. An attack in the United States with aerosolized Y. pestis, which induces the fatal pneumonic form of the disease, would have severe consequences on human lives and the economy. Sporadic outbreaks of the plague also continue to occur naturally in Africa, Asia and the Americas. Strains of Y. pestis that are resistant to multiple antibiotics have recently been isolated, suggesting that natural outbreaks of plague may become more difficult to control. Currently, there are no effective vaccines to protect against Y. pestis, and unless antibiotics are administered immediately after infection, the disease is usually fatal. Research to identify new strategies to control Y. pestis will require genetic manipulation of the pathogen. However, most techniques in current use require the introduction of genetic elements that carry antibiotic resistance markers. Antibiotic resistant Y. pestis poses a health concern not only to researchers, but also as an enhanced risk if resistant strains are obtained by bioterrorists. This research seeks to test the main hypothesis that mutant repressor proteins will function as selectable markers by imparting resistance to potent metabolic inhibitors. These studies will provide new tools for genetic manipulation of Y. pestis that will lead to a better understand its pathogenicity as well as to the production of effective vaccines, without increasing virulence. The methods described here will also be applicable to other select agents of bioterrorism.