Genetic Manipulation of Borrelia burgdorferi
Patricia A. Rosa, Felipe Cabello and D. Scott Samuels
from: Borrelia: Molecular Biology, Host Interaction and Pathogenesis (Edited by: D. Scott Samuels and Justin D. Radolf). Caister Academic Press, U.K. (2010)
Genetic studies in Borrelia burgdorferi require special consideration of the bacterium's segmented genome, complex growth requirements and evolutionary distance from other genetically tractable bacteria, among other features. Despite these challenges, molecular genetics has been applied with growing success to study the biology and virulence of this important human pathogen. In this chapter we summarize the tools and techniques that are currently available for the genetic manipulation of B. burgdorferi, placing 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 B. burgdorferi. We have summarized published methods and highlighted their critical elements, but we have not provided detailed protocols. Although a number of advances have been made since B. burgdorferi was first transformed nearly 15 years ago, some fairly standard components of a good genetic system remain elusive. We mention these limitations as well and why they persist, if known. We hope to encourage investigators to explore what might be possible, in addition to recognizing what currently can be achieved, through genetic manipulation of B. burgdorferi read more ...