Differential Gene Expresion in Yersinia pestis

描述(由申请人提供)：这项建议的长期目标是更好地了解鼠疫杆菌与密切相关的假性结核杆菌(Yptb)区别的分子基础，特别是在其致病能力方面。这一建议解决了这样的假设，即对环境条件的反应的差异基因表达是区分鼠疫杆菌和Yptb的关键决定因素。具体目标是： 目的1：利用双向凝胶电泳法，比较不同生物学条件下鼠疫耶尔森氏菌和Yptb蛋白的表达模式； 目的2：利用质谱仪(MS)鉴定鼠疫耶尔森氏菌对环境的差异表达蛋白的一个子集。这一组将包括鼠疫耶尔森氏菌特异表达的蛋白质，以及一些受鼠疫耶尔森氏菌和Yptb类似调控的蛋白质。MS分析将在以下方面进行： A)从目标1中分析的2D凝胶中分离出的蛋白质； B)差示标记和分级的全细胞蛋白裂解物； 目的：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鉴定，以评估每个基因在未来研究中的潜力。 鼠疫耶尔森氏菌是鼠疫的病原菌，是一种主要的生物威胁因子和A类选择剂。鉴定在生物相关条件下表达的鼠疫杆菌蛋白将为鼠疫检测、治疗和预防提供新的靶点，并深入了解这种至关重要的病原体的发病机制。在未来的研究中，对鼠疫耶尔森氏菌特异基因及其蛋白的表征也将确定鼠疫耶尔森氏菌成为有别于Yptb的新病原体的潜在机制。

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.