UNDERSTANDING THE PATHOGENESIS OF CAMPYLOBACTER JEJUNI

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。我们的长期目标是了解空肠弯曲杆菌的独特生物学和致病性，这是一种重要的人类病原体，也是包括鸡肉在内的食物物种的共生物种，大多数人类病例都来自于空肠弯曲杆菌(疾病控制中心)。我们开发了一些以前无法用于这种微生物研究的基本遗传工具，包括随机和定点突变突变系统、用于研究转录调控的报告质粒和穿梭/表达载体(Hendrixson等人，2001；Hendrixson，O？Rourke和DiRita，未发表)。我们已经使用这些来识别和表征鸡的定植、鞭毛运动、细胞入侵和DNA转化能力所需的基因(Hendrixson等人，2001；Hendrixson和DiRita，未发表，Fogg和DiRita，未发表，Wiesner，Hendrixson和DiRita，正在准备中)。相互作用数据和蛋白质表达谱数据都将有助于我们对空肠弯曲菌的研究。正在进行的不同项目涉及识别和表征蛋白质复合体或评估关键突变体的蛋白质图谱。建议的高通量克隆技术，将单个克隆标记为蛋白质纯化，将对我的团队具有巨大价值，因为它将消除我们生产标记蛋白质以进行个体纯化的需要。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our long objectives are to understand the unique biology and pathogenicity of Campylobacter jejuni, an important human pathogen and commensal of food species including the chicken, from which the majority of human cases originate (Centers for Disease Control). We developed a number of essential genetic tools previously unavailable for the study of this microbe, including random and site specific mutagenesis mutagenesis systems, reporter plasmids for studying transcription regulation and shuttle/expression vectors (Hendrixson, et al., 2001; Hendrixson, O¿Rourke and DiRita, unpublished). We have used these to identify and characterize genes required for colonization of chickens, flagellar motility, cell invasion, and competence for DNA transformation (Hendrixson, et al., 2001; Hendrixson and DiRita, unpublished, Fogg and DiRita, unpublished, Wiesner, Hendrixson and DiRita, in preparation). Interaction data and protein expression profiling data will both be very helpful to our studies on C. jejuni. Different ongoing projects relate to identifying and characterizing protein complexes or assessing the protein profiles of key mutants. High-throughput cloning technology as proposed, leaving the individual clones tagged for protein purification, will be of enormous value to my group as it would obviate the need for us to produce tagged proteins for purification on an individual basis.