Battisti James M, Raffel Sandra J, Schwan Tom G
Division of Biological Sciences, University of Montana, Missoula, MT, USA.
Methods Mol Biol. 2008;431:69-84. doi: 10.1007/978-1-60327-032-8_6.
The lack of a system for genetic manipulation has hindered studies on the molecular pathogenesis of relapsing fever Borrelia. The focus of this chapter is to describe selectable markers, manipulation strategies, and methods to electro-transform and clone wild-type infectious Borrelia hermsii. Preliminary studies suggest that the variable tick protein (Vtp) of B. hermsii is involved in tick-to-mammal transmission. To address this hypothesis, we have developed a system for genetic manipulation and have constructed clones of a Vtp mutant and an isogenic reconstituted strain. The methods described here are applicable for the inactivation of other loci in B. hermsii and should be adaptable for other species of relapsing fever spirochetes.
缺乏基因操作体系阻碍了对复发性发热疏螺旋体分子发病机制的研究。本章重点描述可选择标记、操作策略以及电转化和克隆野生型感染性赫氏疏螺旋体的方法。初步研究表明，赫氏疏螺旋体的可变蜱蛋白（Vtp）参与蜱到哺乳动物的传播。为验证这一假设，我们开发了一种基因操作系统，并构建了Vtp突变体和同基因重组菌株的克隆。此处所述方法适用于赫氏疏螺旋体其他基因座的失活，并且应该适用于其他复发性发热螺旋体物种。
