Differential Gene Expresion in Yersinia pestis

描述（由申请人提供）：本提案的长期目标是更好地了解Y的分子基础。鼠疫菌与近缘种Y.假结核病（Yptb），特别是其致病能力。这一提议解决了这样一个假设，即响应于环境条件的差异基因表达是区分鼠疫杆菌与自然界中的Yptb的关键决定因素。具体目标是： 目的1：比较和对比Y。鼠疫杆菌和Yptb蛋白在多种生物学相关条件下的表达; 目的2：使用质谱（MS）鉴定Y的一个子集。鼠疫菌蛋白质的差异表达，以应对环境。这一组将包括由Y特异性表达的蛋白质。鼠疫菌，以及一些类似的调节，由Y。鼠疫菌和Yptb. MS分析将在以下时间进行： a）从目标1中分析的2D凝胶分离的蛋白质; B）差异标记和分级分离的全细胞蛋白裂解物; 目标3：使用分子和生物信息学方法对目标2中选定的基因进行初步表征，以评估每个基因未来研究的潜力。 鼠疫耶尔森氏菌是鼠疫的病原体，是一种主要的生物威胁因子，也是A类选择因子。鉴定Y.在生物学相关条件下表达的鼠疫蛋白将提供新的靶标，可用于鼠疫检测、治疗和预防，以及深入了解这种至关重要的病原体的发病机制。对Y.在未来的研究中，鼠疫特异性基因及其蛋白质也将确定Y。鼠疫是一种不同于Yptb的新病原体。

DESCRIPTION (provided by applicant): The long-term objective of this proposal is to better understand the molecular basis of Y. pestis differentiation from the closely related species Y. pseudotuberculosis (Yptb), particularly with respect to its ability to cause disease. This proposal addresses the hypothesis that differential gene expression in response to environmental conditions is a critical determinant that distinguishes the plague bacillus from Yptb in nature. The specific aims are to: Aim 1: compare and contrast, using 2D-gel electrophoresis, patterns of Y. pestis and Yptb protein expression in a variety of biologically relevant conditions; Aim 2: identify, using mass spectrometry (MS), a subset of Y. pestis proteins that are differentially expressed in response to the environment. This group will include proteins that are expressed specifically by Y. pestis, as well as some that are similarly regulated by both Y. pestis and Yptb. MS analyses will be done on: a) proteins isolated from 2D gels analyzed in Aim 1; b) differentially-tagged and fractionated whole cell protein lysates; Aim3: perform preliminary characterization of selected genes from Aim 2, using molecular and bioinformatics approaches to evaluate each gene's potential for future study. Yersinia pestis, the etiologic agent of plague, is a major biological threat agent and a Class A select agent. Identification of Y. pestis proteins that are expressed during biologically relevant conditions will provide new targets that can be exploited for plague detection, treatment, and prevention, as well as insights into the pathogenesis of this critically important pathogen. Characterization of Y. pestis-specific genes and their proteins in future studies will also identify potential mechanisms by which Y. pestis emerged as a new pathogen distinct from Yptb.