Purification of Intact Flagella from Escherichia coli and Bacillus subtilis

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 (NH4)2SO4 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.

[1]  J. Adler,et al.  Fine Structure and Isolation of the Hook-Basal Body Complex of Flagella from Escherichia coli and Bacillus subtilis , 1971, Journal of bacteriology.

[2]  M. Simon,et al.  Purification and Thermal Stability of Intact Bacillus subtilis Flagella , 1971, Journal of bacteriology.

[3]  J. Adler,et al.  Attachment of Flagellar Basal Bodies to the Cell Envelope: Specific Attachment to the Outer, Lipopolysaccharide Membrane and the Cytoplasmic Membrane , 1971, Journal of bacteriology.

[4]  M. Simon,et al.  Antigenic Nature of Bacterial Flagellar Hook Structures , 1970, Infection and immunity.

[5]  A. Vatter,et al.  Differentiation Within the Bacterial Flagellum and Isolation of the Proximal Hook , 1970, Journal of bacteriology.

[6]  Z. Vaituzis,et al.  Relationship between Cell Wall, Cytoplasmic Membrane, and Bacterial Motility , 1969, Journal of bacteriology.

[7]  R. Doetsch,et al.  MOTILITY IN PROCARYOTIC ORGANISMS: PROBLEMS, POINTS OF VIEW, AND PERSPECTIVES , 1968, Biological reviews of the Cambridge Philosophical Society.

[8]  M. Simon,et al.  Use of Radioactive Antibodies for Characterizing Antigens and Application to the Study of Flagella Synthesis , 1968, Journal of bacteriology.

[9]  G. Cohen-bazire,et al.  Basal Organelles of Bacterial Flagella , 1967, Journal of bacteriology.

[10]  A. Lawn.  Simple Immunological Labelling Method for Electron Microscopy and its Application to the Study of Filamentous Appendages of Bacteria , 1967, Nature.

[11]  W. van Iterson,et al.  BASAL BODIES OF BACTERIAL FLAGELLA IN PROTEUS MIRABILIS , 1966, The Journal of cell biology.

[12]  F. Dixon,et al.  A method of trace iodination of proteins for immunologic studies. , 1966, International archives of allergy and applied immunology.

[13]  A. Vatter,et al.  Basal Structure and Attachment of Flagella in Cells of Proteus vulgaris , 1965, Journal of bacteriology.

[14]  D. Abram,et al.  IN VITRO FORMATION OF FLAGELLA-LIKE FILAMENTS AND OTHER STRUCTURES FROM FLAGELLIN. , 1964, Journal of molecular biology.

[15]  U. Lewis,et al.  Disk Electrophoresis of Basic Proteins and Peptides on Polyacrylamide Gels , 1962, Nature.

[16]  J. C. Campbell,et al.  The effect of non-lethal deflagellation on bacterial motility and observations on flagellar regeneration. , 1959, Journal of general microbiology.

[17]  R. Repaske,et al.  Lysis of gram-negative organisms and the role of versene. , 1958, Biochimica et biophysica acta.

[18]  H. Epstein,et al.  Optical methods for measuring nucleoprotein and nucleic acid concentrations. , 1957, Archives of biochemistry and biophysics.

[19]  E. H. Melvin,et al.  Determination of Dextran with Anthrone , 1953 .

[20]  Marvin J. Johnson.  ISOLATION AND PROPERTIES OF A PURE YEAST POLYPEPTIDASE , 1941 .

[21]  Dan Hampton Campbell,et al.  Methods in immunology;: A laboratory text for instruction and research , 1970 .