New genetic approaches analyzing Borrelia burgdorferi

描述(由申请人提供)：莱姆病的研究一直受到伯氏疏螺旋体(BB)难治性病原体的阻碍，这种病原体需要进行基因操作。该项目将探索提高基因操作BB效率的新途径；这些进展将用于了解选定基因在宿主(扁虱和哺乳动物)毒力表达和/或定植中的作用(S)。其具体目标是：(1)开发一种新的插入突变系统(称为“RMEM”)，将典型地伴随着Bb的遗传操作的不良的自发毒力丧失降至最低；(2)应用新设计的RMEM突变系统来失活关键调控网络(例如rpoN-rpos)中的基因和那些参与严格类似反应的基因(例如NDK和SPOT)，以阐明它们在调控Bb毒力表达和/或宿主定植中的作用(S)。因此，这项建议寻求为疏螺旋体遗传学贡献重大的新进展，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.