Novel Plasmids for Porphyromonas Post-Genomic Research

产品说明：牙龈卟啉单胞菌被认为是牙周炎的主要病原体，牙周炎是一种在成人中流行的牙齿支撑结构的破坏性疾病。对牙龈卟啉单胞菌的分子遗传学研究已经开始为了解这种生物体如何在口腔中定植并导致病理学奠定基础。随着牙龈卟啉单胞菌W83基因组的核苷酸序列的确定，这种理解可能会急剧增加。到目前为止，这种生物的遗传分析已经可以使用一些从其他细菌系统改编的工具。一些遗传工具已经专门用于卟啉单胞菌，但是如果我们要充分利用该病原体的基因组核苷酸序列，则迫切需要质粒载体的系统构建。我们建议通过开发，测试和分发深入的牙龈卟啉单胞菌后基因组分析所需的质粒构建体来实现这一目标。具体来说，我们将：1。表征来自口腔黑色着色革兰氏阴性厌氧菌的天然和重组质粒在牙龈卟啉单胞菌中稳定支持染色体外复制的能力; 2.设计、构建、表征和测试几种稳定的穿梭载体用于牙龈卟啉单胞菌的分析; 3.使用4种不同的非克隆牙龈卟啉单胞菌菌株，评估可能影响其作为分子遗传分析的宿主的适合性的细胞和分子变量; 4.向研究牙龈卟啉单胞菌和其他黑色素革兰氏阴性厌氧菌的工作人员分发这些新的遗传工具及其使用知识。

DESCRIPTION: Porphyromonas gingivalis is recognized as a prime etiological agent in periodontitis, a destructive disease of the supporting structures of the teeth which is prevalent in adults. Molecular genetic research on P. gingivalis has begun to build a foundation of understanding concerning how this organism colonizes the oral cavity and causes pathology. A dramatic increase in this understanding is likely now that the nucleotide sequence of the genome of P. gingivalis W83 has been determined. To date, genetic analyses of this organism have been possible using a number of tools adapted from other bacterial systems. A few genetic tools have been specifically created for use in Porphyromonas sp., but the systematic construction of plasmid vectors is critically needed if we are to fully exploit the genomic nucleotide sequence of this pathogen. We propose to accomplish this by developing, testing, and distributing plasmid constructs needed for the in-depth postgenomic analyses of Porphyromonas gingivalis. Specifically, we shall: 1. Characterize and native and recombinant plasmids from oral black pigmented Gram negative anaerobes for their ability to stably support extrachromosomal replication in P. gingivalis; 2. Design, build, characterize and test several robust shuttle vectors for the analysis of P. gingivalis; 3. Using 4 different non-clonal P. gingivalis strains, evaluate cellular and molecular variables that may affect their suitability as hosts for molecular genetic analyses; 4. Distribute these new genetic tools and knowledge about their use to workers studying Porphyromonas gingivalis and other black pigmented Gram-negative anaerobes.