Sequence of Rickettsia Rickettsii Genome

描述（由申请人提供）：本提案的具体目标是 测序并注释立克次体的基因组，并开始 基因组的功能分析R.立克次氏体是一种专性细胞内 革兰氏阴性菌，广泛分布于美洲。是 落基山斑疹热的病原体，最严重的人类 立克次体病这种微生物被认为是潜在的生物 武器，因为它的高传染性，因为它会导致疾病与短 潜伏期和相对高的死亡率，特别是在未经治疗的情况下。 目前，对人类的潜在威胁与 健康状况由R.立克次氏体和研究这一问题的实验室数量 重要病原体在现有方法的基础上， 研究这种细菌，这项建议将引入基因组测序作为一个 这意味着获得有价值的深入了解这种复杂的发病机制， 微生物衍生的数据库将为进一步的 功能蛋白质组学研究立克次氏体可能促进 鉴定新的候选疫苗和潜在的治疗靶点。 此外，这些新信息将完善我们对生物学的理解， 这种细菌的进化，也许会导致认识到新的 致病机制的研究。立克次氏体 我们建议完成R. 在一年内完成立克次氏体基因组测序，每对碱基的总成本为0.36美元。一个 初始计算方法将用于识别假定开放阅读 编码蛋白质和表达为RNA的基因的ORF。一个新 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 成分在R.立克次氏体将通过计算分析 操纵子的结构和比较蛋白质和途径，在R。普劳瓦泽基 和其他细菌。为了便于实验功能基因组分析， R.立克次氏体，将单个ORF进行PCR扩增、克隆、表达， 编目并提供给科学界。生化 对R.将通过实验证实立克次氏体。 最后，选择了转录调控子。立克次氏体， 将表征外部信号。

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.