Pathway for functional characterization of hypothetical genes and non-coding RNAs of Enterococcus faecalis

 描述(由申请人提供)：本项目将研究粪肠球菌非编码调控RNA(NcRNAs)和假想蛋白在适应与机会性感染相关的生长条件方面的作用。粪肠球菌是医院获得性感染的主要原因，其治疗受到有机体固有和后天的抗生素耐药性、适应恶劣条件下的生长和生存的倾向以及在共生和致病生活方式之间转换的能力的阻碍。我们将在体外生物膜系统(CDC生物膜反应器)和实验性心内膜炎模型中研究实验室模型菌株OG1RF。我们将使用计算搜索和一种功能强大的新遗传屏幕相结合的方式来识别在这些环境中有助于竞争适应性的假设蛋白质ncRNA，并开始研究它们的目标和作用机制。这项研究的结果将包括：1)将出现一个更全面的图景，在生长条件下，与其在医院感染中的作用相关的ncRNA调节和假设的蛋白质功能在该生物体中的作用。2)将验证粪肠球菌ncRNAs和假想蛋白质调控的全基因组功能和机制研究的工作流程。3)从长远来看，可能会确定新的药物和疫苗目标。

 DESCRIPTION (provided by applicant): This project will examine the role of non-coding regulatory RNAs (ncRNAs) and hypothetical proteins of Enterococcus faecalis in adaptation to growth conditions relevant to opportunistic infections. E. faecalis is a leading cause of hospital-acquired infections whose treatment is impeded by the inherent and acquired antibiotic resistance of the organism, and by its propensity to adapt to growth and survival in hostile conditions, and by its ability to transition between commensal and pathogenic lifestyles. We will study the model laboratory strain OG1RF in both an in-vitro biofilm system (CDC biofilm reactor) and in an experimental endocarditis model. We will use a combination of computational searches, and a powerful new genetic screen we have termed "Smart TnSeq" to identify hypothetical proteins ncRNAs contributing to competitive fitness in these environments, and begin to study their targets and mechanisms of action. The results of this research will include the following: 1) A more comprehensive picture of the role of ncRNA regulation and hypothetical protein function in this organism under growth conditions relevant to its role in hospital infectios will emerge. 2) A workflow for genome-wide functional and mechanistic studies of regulation by E. faecalis ncRNAs and hypothetical proteins will be validated. 3) In the longer term, new drug and vaccine targets may be identified.