Purification of Intact Flagella from Escherichia coli and Bacillus subtilis

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Abstract
A procedure is described for the purification of bacterial flagella in the form of a filament-hook-basal body complex (intact flagella) free from detectable cell wall, membrane, or cytoplasmic material. Spheroplasts produced with lysozyme and ethylenediaminetetraacetic acid were lysed with Triton X-100, and the flagella were purified by (NH 4 ) 2 SO 4 precipitation, differential centrifugation, and CsCl gradient centrifugation. As much as 40% of the flagella were recovered, and they contained about one basal body per 4 to 6 μm of flagella. The same procedure developed for Escherichia coli was also successful for purifying intact flagella from Bacillus subtilis .