GENETIC APPROACH TO MYCOPLASMA PNEUMONIA VIRULENCE

无细胞壁细菌柔膜菌属（Mollicutes）的成员属于 人类所知的最小和最简单的原核生物。它们拥有非常小的 基因组（500-1000兆道尔顿）和有限的生物合成能力， 因此只能过着寄生或寄生生活。物种 肺炎支原体是人类呼吸道的病原体， 是导致大龄儿童和年轻人肺炎的主要原因。作为 我对这种微生物非常着迷， 发病机制的研究。我通过我的 博士后培训，以熟悉分子生物学的工具， 生物学家，预计将分子克隆应用于研究 发病机制我熟悉从M. pneumoniae用于大肠埃希菌的研究，并使用这种方法 研究支原体粘附宿主细胞时相变化。但 从这些研究中获得的信息将得到加强 如果克隆与毒力相关的支原体基因 （特别是细胞吸附和氧自由基的产生）可以突变， 回到支原体染色体上， 许多细菌病原体。我的目标是利用这个RCDA 在基因工程方面扩展我的技能， 和M.肺炎。该奖项将使 让我专注于实验问题，并花时间与 和埃默里大学附近的微生物遗传学家一起工作 我可以充分利用同源重组策略。这 部门传统上在分子方法方面有很强的基础， 微生物生理学和遗传学，所描述的研究将允许 扩大基础以包括微生物发病机理。

Members of the class of cell wall-less bacteria Mollicutes are among the smallest and simplest prokaryotes known to man. They possess very small genomes (500-1000 megadaltons) and limited biosynthetic capabilities and hence resort to either a commensal or parasitic lifestyle. The species Mycoplasma pneumoniae is a pathogen of the human respiratory tract and the leading cause of pneumonia among older children and young adults. As a graduate student I became fascinated by this microbe and focused my research on the mechanisms of pathogenesis. I sought through my post-doctoral training to become acquainted with the tools of the molecular biologist, anticipating applying molecular cloning to the study of pathogenesis. I am familiar with techniques for taking genes out of M. pneumoniae for study in Escherichia coli and am using this approach to study phase variation in mycoplasma adherence to host cells. However, the harvest of information to be gained from those studies would be enhanced immensely if cloned mycoplasma genes associated with virulence (specifically cytadsorption and oxyradical generation) could be mutated and returned to the mycoplasma chromosome in the same manner as is now common with many enbacterial pathogens. It is my goal to utilize this RCDA to expand my repertoire in genetic engineering in developing a transforation and cassette mutagenesis system for M. pneumoniae. This award would enable me to focus on the experiment problem and to spend the time interacting with microbial geneticists on this camp and nearby at Emory University so that I might make the most of homologous recombination strategies. This department has traditionally had a strong base in molecular approaches to microbial physiology and genetics, and the studies described would allow expansion of that base to include microbial pathogenesis.