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、肺鼠疫和败血症鼠疫的病原体，目前作为一种潜在的生物恐怖主义武器而备受关注。在美国发生的一起用雾化的Y染色体的袭击。鼠疫会引致致命的肺炎，对人命和经济造成严重后果。在非洲、亚洲和美洲，鼠疫的零星爆发也继续自然发生。菌株Y.对多种抗生素具有抗药性的鼠疫最近被分离出来，这表明鼠疫的自然爆发可能变得更加难以控制。目前，还没有有效的疫苗来预防Y。除非在感染后立即使用抗生素，否则这种疾病通常是致命的。研究确定新的策略来控制Y。鼠疫需要对病原体进行基因操作。然而，目前使用的大多数技术需要引入携带抗生素抗性标记的遗传元件。抗生素抗性Y.鼠疫不仅对研究人员构成健康问题，而且如果生物恐怖分子获得耐药菌株，风险也会增加。本研究旨在验证突变阻遏蛋白通过赋予对有效代谢抑制剂的抗性而作为选择标记发挥作用的主要假设。这些研究将为Y染色体的遗传操作提供新的工具。这将导致更好地了解其致病性以及生产有效的疫苗，而不增加毒性。这里所述的方法也适用于其他生物恐怖主义的选择剂。

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.