Purification of Fla2 flagella of Rhodobacter sphaeroides

Abstract

The photosynthetic bacterium R. sphaeroides expresses two flagellar systems that are encoded by two complete gene clusters that have distinct phylogenetic origins. The isolation and purification of the Filament-Hook Basal Body (F-HBB) or the Hook Basal Body (HBB) structure is a troublesome task given the complexity of this nano-machine that is composed of multiple loosely bound substructures that can be lost during the isolation and purification procedure. A successful procedure requires adjustments to the standard method established for Salmonella. In this chapter, we describe a detailed protocol to isolate and purify the Fla2 F-HBB and HBB from R. sphaeroides a photosynthetic bacterium that has a complex intracellular membrane system that frequently interferes with isolation of high-quality samples.