PORPHYROMONAS GINGIVALIS GENOME PROJECT

描述（改编自研究者摘要）：长期 该研究的目的是开发一种全面的分子生物学方法， 了解有机体牙龈卟啉单胞菌，一个物种强烈 与人类牙周病有关。 牙龈卟啉单胞菌是一个小基因组 噬纤维菌/黄细菌/拟杆菌的代表性研究较少 门，一组重要的生物体。 的 本研究将获得强毒株W83的全基因组序列，约2.2Mb， 目标1（Aim 1） 对于测序，使用2.0 kb平均插入片段大小的文库 将在pUC 18载体和18. kb平均插入片段大小 lambda DashII中的函数库。 大约35，200个序列来自于 17，600个pUC克隆将产生约8倍的基因组覆盖率。 还将对1，000个lambda DashII克隆的末端进行测序，以创建 基因组支架，并验证重叠群的方向和完整性 由来自质粒文库的序列的组装形成。 基因组 序列将从随机测序的 片段;测序缺口将通过步移适当的克隆来闭合，并且 通过聚合酶链反应的物理间隙。 我们会分析这个序列 注（目标2）。 将识别开放阅读框架（Orfs） 与已知和未知功能的蛋白质有关。 序列将是 分析以确定许多特征，包括代谢途径， 毒力因子和输出蛋白。 DNA结构基序（如 启动子、复制起点和重复元件）和rRNA的基因，以及 将鉴定小RNA，如tRNA、4.5S、6S、10 S和RNAse P。 来自注释序列的信息将与 来自牙龈卟啉单胞菌研究社区的实验数据，包括 体内表达技术（目的3）。 基因组序列和注释数据 将在TIGR微生物数据库网站上提供，以及 保藏在GenBank和其他适当的数据库中（目标4）。 序列 本提案中产生的信息将加速正在进行的牙龈卟啉单胞菌 研究，以确定和表征毒力因子，并制定一个 有效的牙龈卟啉单胞菌疫苗 五分之一的人患有牙周病 因此，预防和治疗方面的进展将对美国人产生深远的影响。 对美国人民的健康产生重大影响， 牙齿护理。

DESCRIPTION (Adapted from the Investigator's Abstract): The long-term objective of the proposed research is to develop a comprehensive molecular understanding of the organism Porphyromonas gingivalis, a species strongly associated with human periodontal diseases. P. gingivalis is a small genome representative of the little studied Cytophaga/Flavobacteria/Bacteroides phylum, a phylogenetically and medically important group of organisms. The complete genome sequence of virulent strain W83, about 2.2 Mb, will be determined (Aim 1). For sequencing, a 2.0 kb average insert size library will be prepared in the pUC18 vector and an 18. kb average insert size library in lambda DashII. Approximately 35,200 sequences from both ends of 17,600 pUC clones will yield approximately 8-fold coverage of the genome. The ends of 1,000 lambda DashII clones will also be sequenced to create a genome scaffold, and to verify the orientation and integrity of the contigs formed from the assembly of sequences from the plasmid library. The genome sequence will be assembled from the collection of randomly sequenced fragments; sequencing gaps will be closed by walking appropriate clones, and physical gaps by polymerase chain reaction. The sequence will be analyzed and annotated (Aim 2). Open reading frames (Orfs) will be identified related to proteins of known and unknown function. The sequence will be analyzed to identify a number of features, including metabolic pathways, virulence factors, and exported proteins. DNA structural motifs (such as promoters, replication origins and repeat elements) and genes for rRNA, and small RNAs, such as tRNA, 4.5S, 6S, 10S, and RNAse P, will be identified. The information from the annotated sequence will be integrated with experimental data from the P. gingivalis research community, including in vivo expression technology (Aim 3). The genome sequence and annotation data will be made available on the TIGR Microbial Database web site, as well as deposited in GenBank and other appropriate databases (Aim 4). The sequence information generated in this proposal will accelerate ongoing P. gingivalis research to identify and characterize virulence factors and to develop an effective P. gingivalis vaccine. Periodontal disease affects one in five Americans, and therefore, advances in prevention and therapy will have a significant effect on the health of the American people, and the cost of dental care.