Proteomics study of Bacteroides forsythus

描述：病原微生物通过表达称为毒力因子的特定基因产物在宿主内生存并造成损害。尽管越来越多的证据表明福赛斯拟杆菌是一种重要的牙周病细菌，但由于对其毒力因子的了解有限，这种微生物启动和介导致病过程的确切机制几乎是未知的。因此，为了提高对Bf感染的认识，有必要对Bf未知的毒力因子进行鉴定和表征。 本申请提供的初步数据表明，比较双向凝胶电泳法(2-DE)可用于检测Bf的差异表达蛋白质，从而发现体内选择性表达的蛋白质。本应用的目的是利用蛋白质组学研究鉴定和鉴定体内表达的Bf蛋白。预计这种新的方法与Bf基因组序列的分析相结合，将导致对Bf潜在毒力因子的检测、鉴定和表征。 为了实现这一目标，提出了以下具体目标：1)利用双向凝胶电泳法检测Bf的差异表达蛋白。通过2-DE比较体外和体内生长的Bf细胞的蛋白质谱，将检测到体内选择性表达的蛋白质；2)利用质谱学和数据库搜索来鉴定和鉴定差异表达的蛋白质。体内表达的蛋白质将通过质谱学和搜索Bf基因组和蛋白质数据库来确定。这种方法将为产生新的假说提供基础，这些假说可以在未来的研究中检验，在这些假说中，候选毒力因子的真实毒力函数可以得到验证。这些研究的结果将有助于实现本实验室的长期目标：通过提供更多关于BF毒力因子的信息，更好地了解BF的致病机理。

DESCRIPTION: Pathogenic microorganisms survive and cause damage within the host by expressing specific gene products called virulence factors. Despite emerging evidences indicating that Bacteroides forsythus (Bf) is a crucial periodontopathic bacterium, the exact mechanisms whereby this microorganism initiates and mediates the pathogenic process are virtually unknown because of the limited information on its virulence factors. Therefore, in order to advance understanding of Bf infection, it is required to identify and characterize unknown virulence factors of Bf. Preliminary data presented in this application indicated that comparative 2-dimensional gel electrophoresis (2-DE) could be used to detect differentially expressed proteins of Bf, and the proteins selectively expressed in vivo were thus discovered. The purpose of this application is to identify and characterize in vivo-expressed proteins of Bf using proteomics study. It is expected that this novel approach, combined with the analysis of the Bf genome sequence, will lead to detection, identification, and characterization of potential virulence factors of Bf. In order to achieve this goal, the following Specific Aims are proposed: 1) to detect differentially expressed proteins of Bf by 2-DE. By comparing the protein profiles of in vitro- and in vivo-grown Bf cells using 2-DE, the proteins selectively expressed in vivo will be detected; 2) to Identify and characterize differentially expressed proteins using mass spectrometry and database search. Identity of in vivo-expressed proteins will be determined by mass spectrometry and search for Bf genome and protein database. This approach will provide a foundation for generating novel hypotheses that can be tested in future studies, in which authentic virulence function of the candidate virulence factors can be validated. The results obtained from these studies will assist to achieve a long-term goal of this laboratory: to gain a better understanding of the pathgogenic of Bf by providing more information on its virulence factors.