Sequence of Rickettsia Rickettsii Genome

描述(由申请人提供)：本提案的具体目的是 对立克次体基因组进行测序和注释，并开始 基因组的功能分析。立克次体是一种细胞内必备的 广泛分布在美洲的革兰氏阴性细菌。它是 落基山斑疹热的病原体是人类最严重的 立克次体病。这种微生物被认为是一种潜在的生物 武器因为它的高传染性，因为它引起疾病与短的 潜伏期和相对较高的死亡率，特别是在未经治疗的病例中。 目前，对人类的潜在威胁之间存在严重的分歧 立克次体构成的健康以及研究这一问题的实验室数量 重要的病原体。添加到目前可用的现有方法中 对这种细菌的研究，这项提议将引入基因组测序作为一种 手段，以获得宝贵的洞察力的错综复杂的发病机制 微生物。派生的数据库将为进一步的开发奠定基础 立克次体功能蛋白质组学研究可能有助于 确定新的候选疫苗和潜在的治疗靶点。 此外，这些新信息将加深我们对生物学的理解。 这种细菌的进化，也许会导致对新奇事物的认识 立克次体致病机制的研究。 建议完成R. 一年内获得立克次体基因组，每对碱基对的总成本为0.36美元。一个 初步的计算方法将用于确定假定的开放阅读 编码以RNA形式表达的蛋白质和基因的框架(ORF)。一个新的 开发的网站将建设和维护，以显示R。 立克次体基因组序列及其注释。代谢途径及其研究进展 立克次体中的成分将通过计算分析确定 普氏乳杆菌操纵子结构及其与蛋白质和通路的比较 和其他细菌。为便于进行实验功能基因组分析 利用聚合酶链式反应扩增、克隆、表达单个开放阅读框， 被编目，并提供给科学界。生化 立克次体选定基因的功能将得到实验证实。 最后，筛选出立克次体的转录调控因子。 将对外部信号进行表征。

DESCRIPTION (provided by applicant): The specific aims of this proposal are to sequence and annotate the genome of Rickettsia rickettsii, and to begin functional analysis of the genome. R. rickettsii is an obligately intracellular Gram negative bacterium that is widely distributed in the Americas. It is the etiologic agent of Rocky Mountain spotted fever, the most severe of the human rickettsioses. This microorganism has been considered as a potential biological weapon because of its high infectivity, since it causes disease with a short incubation period and relatively high mortality, especially in untreated cases. Currently, a serious discrepancy exists between the potential threat to human health posed by R. rickettsii and the number of laboratories studying this important pathogen. To add to existing methodologies presently available for study of this bacterium, this proposal will introduce genomic sequencing as a means to gain valuable insight into the intricacies of the pathogenesis of this microorganism. The derived database will lay the groundwork for further functional proteomic studies of R. rickettsii that may facilitate identification of new vaccine candidates and potential therapeutic targets. Additionally, this new information will refine our understanding of the biology of the evolution of this bacterium and perhaps result in recognition of novel pathogenic mechanisms used by R. rickettsii. We propose to complete the sequencing and primary annotation of the R. rickettsii genome within one year at a total cost of $0.36 per base pair. An initial computational approach will be used to identify putative open reading frames (ORF) encoding the proteins and genes expressed as RNAs. A newly developed Website will be constructed and maintained to display the R. rickettsii genome sequence and its annotation. Metabolic pathways and their components in R. rickettsii will be identified by computational analysis of operon structures and comparisons with proteins and pathways in R. prowazekii and other bacteria. To facilitate experimental functional genomic analyses of R. rickettsii, individual ORFs will be PCR amplified, cloned, expressed, catalogued, and made available to the scientific community. Biochemical functions of selected genes of R. rickettsii will be experimentally confirmed. Finally, selected transcriptional regulons of R. rickettsii which respond to external signals will be characterized.