Genetic studies in Borrelia require special consideration of the highly segmented genome, complex growth requirements and evolutionary distance of spirochetes from other genetically tractable bacteria. Despite these challenges, a robust molecular genetic toolbox has been constructed to investigate the biology and pathogenic potential of these important human pathogens. In this review we summarize the tools and techniques that are currently available for the genetic manipulation of Borrelia, including the relapsing fever spirochetes, viewing them in the context of their utility and shortcomings. Our primary objective is to help researchers discern what is feasible and what is not practical when thinking about potential genetic experiments in Borrelia. We have summarized published methods and highlighted their critical elements, but we are not providing detailed protocols. Although many advances have been made since B. burgdorferi was first transformed over 25 years ago, some standard genetic tools remain elusive for Borrelia. We mention these limitations and why they persist, if known. We hope to encourage investigators to explore what might be possible, in addition to optimizing what currently can be achieved, through genetic manipulation of Borrelia.

中文翻译：

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疏螺旋体的遗传操作。

疏螺旋体的遗传研究需要特别考虑高度分段的基因组、复杂的生长要求以及螺旋体与其他遗传易处理细菌的进化距离。尽管存在这些挑战，但已经构建了一个强大的分子遗传工具箱来研究这些重要人类病原体的生物学和致病潜力。在这篇综述中，我们总结了目前可用于对疏螺旋体进行基因操作的工具和技术，包括回归热螺旋体，并从它们的实用性和缺点的角度来看待它们。我们的主要目标是帮助研究人员在考虑潜在的疏螺旋体基因实验时辨别什么是可行的，什么是不实用的. 我们总结了已发布的方法并强调了它们的关键要素，但我们没有提供详细的协议。尽管自25 年前首次转化伯氏疏螺旋体以来已经取得了许多进展，但对于疏螺旋体而言，一些标准的遗传工具仍然难以捉摸。我们提到了这些限制以及为什么它们会持续存在（如果知道的话）。我们希望鼓励研究人员通过对疏螺旋体的基因操作来探索什么是可能的，除了优化目前可以实现的目标。