An analysis of Borrelia burgdorferi motility

描述（由申请人提供）：莱姆病是美国最普遍的节肢动物传播的感染，自发现以来，发病率稳步上升。螺旋体伯氏疏螺旋体是病原体。对B的了解很少。将疾病带给宿主的伯氏菌毒力属性。目前的建议集中在B的趋化性和运动性。burgdorferi，以及它们在疾病过程中的作用。趋化性和运动性可能是重要的毒力属性，因为这些生物体渗透到其他细菌入侵的组织和器官中。为了在分子水平上理解趋化性，在目标1中，我们将确定那些作为引诱剂的化合物。推定的化学引诱物将根据从基因组序列和B的已知代谢中提取的信息来鉴定。这些化合物的趋化能力将使用已建立的方法进行测试。将在适当的环境条件下检测高传代（无毒力）和低传代（有毒力）菌株。在目标2中，我们提出在特定运动和趋化基因中构建插入缺失和框内缺失突变。高传代和低传代菌株均用作亲本菌株。我们假设这些突变体中的每一个将具有不同的表型。将使用DNA微阵列技术分析这些表型的运动性、趋化性、结构、蛋白质印迹、蛋白质组学和转录分析。为了确保一个给定的表型是由于正在研究的靶向突变，我们将使用基因互补分析。在目标3中，我们将检验运动性和趋化性是B的重要毒力属性的假设。burgdorferi。为了验证这一假设，我们将使用标准的细胞渗透试验和莱姆病的小鼠模型。我们预测，运动和趋化性突变体将是较低的侵入性和毒性比亲本菌株在这两个测定。所提出的实验应该产生关于B的趋化性和运动性的作用的信息。莱姆病中的burgdorferi，并可能为预防和治疗的新方法奠定基础。

DESCRIPTION (provided by applicant): Lyme disease is the most prevalent arthropod borne infection in the United States, and has shown a steady increase in incidence since its discovery. The spirochete Borrelia burgdorferi is the causative agent. Very little is known about the B. burgdorferi virulence attributes that bring disease to the host. The present proposal focuses on the chemotaxis and motility of B. burgdorferi, and their rote in the disease process. Chemotaxis and motility are likely to be important virulence attributes, as these organisms penetrate into tissues and organs where other bacteria fall to invade. Basic to an understanding of chemotaxis on a molecular level, in Aim 1 we will identify those compounds that serve as attractants. Putative chemoattractants will be identified from information drawn from the genome sequence and known metabolism of B. burgdorferi The chemotactic ability of these compounds will be tested using established methodology. Both high passage (avirulent) and tow passage (virulent) strains will be tested under appropriate environmental conditions. in Aim 2, we propose to construct both insertion-deletion and in-frame deletions mutations in specific motility and chemotaxis genes. Both high and low passage strains will serve as parental strains. We hypothesize that each of these mutants will have a distinct phenotype. These phenotypes will be analyzed with respect to motility, chemotaxis, structure, western blot, proteomics and transcriptional analysis using DNA-microarray technology. To insure that a given phenotype is due to the targeted mutation being studied, we will use gene-complementation analysis. in Aim 3, we will test the hypothesis that motility and chemotaxis are important virulence attributes of B. burgdorferi. To test this hypothesis, we will use standard cell penetration assays, and the mouse model of Lyme disease. We predict that the motility and chemotaxis mutants will be less invasive and virulent than the parental strains in both of these assays. The experiments proposed should yield information with respect to the roles of chemotaxis and motility of B. burgdorferi in Lyme disease, and perhaps lay the foundation for new methods for prevention and treatment.