Interactions Between Oral Pathogens and Vascular Cells

描述(申请人提供)：牙龈卟啉单胞菌是成人牙周炎的主要病原体。虽然已被公认为牙周病原体，但最近人们认识到牙龈假单胞菌可能在系统性疾病中发挥重要作用，包括心血管疾病、早产和低出生体重。心血管疾病是一组不同的疾病，在发达国家是主要的死亡原因。因此，这种病原体造成的经济损失和心理影响可能是巨大的。在过去的几年里，我们研究了牙龈假单胞菌的致病特性，重点是它与包括冠状动脉内皮细胞在内的人体组织的相互作用和侵袭。然而，仅有一株牙龈假单胞菌的全基因组序列的可用性阻碍了我们的研究努力。许多因素表明，由于菌株在某些毒力因素包括相对宿主细胞侵袭能力方面是表型和基因异质性的，因此应该继续对其他牙龈假单胞菌菌株进行调查。因此，本意见书要求提供资金，作为我们最近资助的竞争性更新(DE13545)的补充，以对牙龈假单胞菌381、33277、A7436和AJW4菌株的基因组进行测序和注释，然后进行比较基因组分析。基因组序列将使用454生命科学公司(康涅狄格州布兰福德)开发的一种新的、具有成本效益的大规模并行DMA测序技术来确定。基因组组装将补充高达1倍的基因组覆盖使用配对的末端桑格DNA测序读数。基因位置和结构将使用FGENESB进行预测。基因功能和名称将被分配给基于公共基因数据的同源搜索而预测的基因结构。将进行成对的基因组比较和多个基因组的比较，以确定在这五个基因组中保守的同源基因以及独特的基因。这些基因组中缺失的基因也将被确定。BLAST搜索、木乃伊程序和Vista工具将用于这些目的。对于所有预测的蛋白质，将进行HMMER PFAM搜索以检测功能结构域。在公共数据库中提供另外四个牙龈假单胞菌的全基因组序列将有助于基础研究，包括为这一补充应用提供的父母资助，以及更快地开发诊断测试、治疗和预防策略。

DESCRIPTION (provided by applicant): Porphyromonas gingivalis is the major etiological agent of adult periodontitis. Although well established as a periodontal pathogen, it has more recently been recognized that P. gingivalis may have an important role in systemic disease, including cardiovascular diseases, prematurity and low birth weight. Cardiovascular diseases are a heterogeneous group of conditions that are the leading cause of death in developed countries. The economic losses and psychological repercussions caused by this pathogen are thus potentially enormous. During the past several years, we have investigated the pathogenic properties of P. gingivalis, focusing on its interactions with and invasion of human tissues, including coronary artery endothelial cells. However, the availability of the whole genome sequence of only one strain of P. gingivalis has hampered our research effort. Many factors suggest that investigation of other P. gingivalis strains should be pursued since strains are phenotypically and genotypically heterogeneous with regards to certain virulence factors including relative host cell invasive abilities. Consequently, this submission is requesting funding as a supplement to our recently funded competitive renewal (DE13545) to sequence and annotate the genomes of P. gingivalis strains 381, 33277, A7436, and AJW4, followed by comparative genomic analysis. Genome sequences will be determined using a new, cost-efficient, and massively parallel DMA sequencing technology developed by 454 Life Sciences (Branford, CT). Genome assembly will be supplemented with up to 1x genome coverage using paired-end Sanger DNA sequencing reads. Gene location and structures will be predicted using FGENESB. Gene function and names will be assigned to predicted gene structures based on homology searches of public gene data. Pairwise genome comparisons and multiple genome comparisons will be performed to identify orthologs that are conserved among these five genomes as well as unique genes. Genes that are missing from these genomes will also be determined. BLAST searches, MUMmer programs and VISTA tools will be used for these purposes. For all predicted proteins, a HMMER PFAM search will be conducted to detect functional domains. The availability of four additional whole genome sequences of P. gingivalis in public databases would facilitate basic research, including that of the parent grant to this supplemental application, and more rapid development of diagnostic tests, and treatment, and prevention strategies.