DEVELOPMENT OF A GENETIC SYSTEM FOR BORRELIA BURGDORFERI

描述（改编自申请人的摘要）：莱姆病，最 在美国，一种常见的节肢动物传播疾病，由感染引起 伯氏疏螺旋体 缺乏一个系统， 从基因上操纵B。burgdorferi阻碍了研究其 分子遗传水平上的生物学和发病机理。 若干组件 需要创建一个有用的遗传系统，用于测定 个体基因 这些包括选择标记，稳定的质粒载体， 和转化方法。 该项目旨在开发一种基因 B系统。burgdorferi，最初基于电穿孔和抗性 抗生素香豆霉素A1 这支笔将用于 通过插入和选择基因来维持广泛的宿主范围 载体和重组天然质粒。 此外，其他可选 将筛选用于疏螺旋体系统的遗传标记。 建立这样一个系统将有助于确定基因缺陷， 突变株的互补，将加快质粒的研究 复制的 不同抗生素载体的构建 抗性标记将允许基因在体外失活， 体内野生型基因座的替换，以及 缺损 这些遗传技术将是系统地 剖析生长和发病所需的基因。

DESCRIPTION (Adapted from the applicant's abstract): Lyme disease, the most common arthropod-borne disease in the United States, is caused by infection with the spirochete Borrelia burgdorferi. The lack of a system to genetically manipulate B. burgdorferi has hindered attempts to study its biology and pathogenesis on a molecular genetic level. Several components are required to create a useful genetic system for assaying the effect of individual genes. These include selectable markers, stable plasmid vectors, and a method of transformation. This project proposes to develop a genetic system for B. burgdorferi, initially based on electroporation and resistance to the antibiotic coumermycin A1. This marker will be used to inactivate genes by insertion and to select for the maintenance of broad host range vectors and recombinant natural plasmids. In addition, other selectable genetic markers will be screened for use in the borrelia system. Establishment of such a system will facilitate identifying gene defects in mutant strains by complementation and will expedite the study of plasmid replication. Construction of several vectors with different antibiotic resistance markers will permit the inactivation of genes in vitro, replacement of the wild type locus in vivo, and complementation of the defect. These genetic techniques will be the first steps in systematically dissecting the genes that are required for growth and pathogenesis.