New genetic approaches analyzing Borrelia burgdorferi

描述（由申请人提供）：莱姆病的研究一直受到病原体伯氏疏螺旋体（Bb）难以进行基因操作的阻碍。该项目将探索新的途径，以提高遗传操纵Bb的效率;这些发展将被应用于理解所选基因在宿主（蜱和哺乳动物）的毒力表达和/或定殖中的作用。具体目标是：（1）开发一种新的插入诱变系统（称为“RMEM”），其将使通常伴随Bb的遗传操作的不期望的毒力自发丧失最小化，和（2）将新设计的RMEM诱变系统应用于关键调控网络中的突变基因（例如，rpoN-rpoS）和参与类似严格反应的那些（例如ndk和spoT），以阐明它们在调节Bb毒力表达和/或宿主定殖中的作用。因此，该提案旨在为疏螺旋体遗传学的重大新进展做出贡献，并利用这些进展来研究Bb毒力表达的潜在新方面。

DESCRIPTION (provided by applicant): The study of Lyme disease has been hampered by the intractability of the etiological agent, Borrelia burgdorferi (Bb), to be genetically manipulated. This project will explore new avenues to improve the efficiency of genetically manipulating Bb; such developments will be applied towards understanding the role(s) of selected genes in virulence expression and/or colonization of hosts (ticks and mammals). The Specific Aims are: (1) To develop a novel insertion mutagenesis system (termed "RMEM") that will minimize the undesirable spontaneous loss of virulence that typically accompanies the genetic manipulation of Bb and (2) To apply the newly designed RMEM mutagenesis system to inactivate genes in a key regulatory network (e.g., rpoN-rpoS) and those involved in a stringent-like response (e.g. ndk, and spoT) to clarify their role(s) in regulating virulence expression and/or host colonization by Bb. This proposal thus seeks to contribute major new advances to borrelial genetics and to use those developments to examine potentially novel aspects of Bb virulence expression.