Purification of intact flagella from Escherichia coli and Bacillus subtilis.

Journal of bacteriology ·Vol. 105 ·No. 1 ·1971-01-00 ·Pages 376-83

DePamphilis ML, Adler J

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 mum of flagella. The same procedure developed for Escherichia coli was also successful for purifying intact flagella from Bacillus subtilis.

MeSH Terms

Bacillus subtilis/analysis,cytology,growth & development Bacterial Proteins/analysis Carbohydrates/analysis Centrifugation, Density Gradient Cesium Chlorides Detergents Dialysis Quaternary Ammonium Compounds

Chemicals

Bacterial Proteins Carbohydrates Chlorides Detergents Quaternary Ammonium Compounds Cesium

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

DePamphilis M L

Adler J

References (21)

21 references, click to expand

Relationship between cell wall, cytoplasmic membrane, and bacterial motility.
J Bacteriol. 1969 Oct;100(1):512-21 PMID: 5344110
A method of trace iodination of proteins for immunologic studies.
Int Arch Allergy Appl Immunol. 1966;29(2):185-9 PMID: 4160044
DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS.
Ann N Y Acad Sci. 1964 Dec 28;121:404-27 PMID: 14240539
The effect of non-lethal deflagellation on bacterial motility and observations on flagellar regeneration.
J Gen Microbiol. 1959 Jun;20(3):670-85 PMID: 13664913
Motility in procaryotic organisms: problems, points of view, and perspectives.
Biol Rev Camb Philos Soc. 1968 Aug;43(3):317-62 PMID: 4920212
Basal organelles of bacterial flagella.
J Bacteriol. 1967 Aug;94(2):458-65 PMID: 6039362
Basal structure and attachment of flagella in cells of Proteus vulgaris.
J Bacteriol. 1965 Nov;90(5):1337-54 PMID: 5848332
Basal bodies of bacterial flagella in Proteus mirabilis. I. Electron microscopy of sectioned material.
J Cell Biol. 1966 Dec;31(3):585-601 PMID: 5971649
Disk electrophoresis of basic proteins and peptides on polyacrylamide gels.
Nature. 1962 Jul 21;195:281-3 PMID: 14491328
The structure of flagella and the genetic control of flagellation in Eubacteriales. A review.
Antonie Van Leeuwenhoek. 1968;34(2):205-25 PMID: 4875403
Lysis of gram-negative organisms and the role of versene.
Biochim Biophys Acta. 1958 Nov;30(2):225-32 PMID: 13607436
Antigenic nature of bacterial flagellar hook structures.
Infect Immun. 1970 Feb;1(2):212-3 PMID: 16557718
Attachment of flagellar basal bodies to the cell envelope: specific attachment to the outer, lipopolysaccharide membrane and the cyoplasmic membrane.
J Bacteriol. 1971 Jan;105(1):396-407 PMID: 4250610
Simple immunological labelling method for electron microscopy and its application to the study of filamentous appendages of bacteria.
Nature. 1967 Jun 10;214(5093):1151-2 PMID: 4861237
IN VITRO FORMATION OF FLAGELLA-LIKE FILAMENTS AND OTHER STRUCTURES FROM FLAGELLIN.
J Mol Biol. 1964 Jul;9:168-85 PMID: 14200380
Purification and thermal stability of intact Bacillus subtilis flagella.
J Bacteriol. 1971 Jan;105(1):369-75 PMID: 4993323
Genetics of motility in Escherichia coli: complementation of paralysed mutants.
Genetics. 1967 Jul;56(3):363-73 PMID: 4861166
Optical methods for measuring nucleoprotein and nucleic acid concentrations.
Arch Biochem Biophys. 1957 May;68(1):144-9 PMID: 13435902
Differentiation within the bacterial flagellum and isolation of the proximal hook.
J Bacteriol. 1970 Jan;101(1):250-61 PMID: 4189228
Use of radioactive antibodies for characterizing antigens and application to the study of flagella synthesis.
J Bacteriol. 1968 Jan;95(1):81-6 PMID: 4965981
Fine structure and isolation of the hook-basal body complex of flagella from Escherichia coli and Bacillus subtilis.
J Bacteriol. 1971 Jan;105(1):384-95 PMID: 4993325

Article Info

Journal

Journal of bacteriology

Abbr.

J Bacteriol

ISSN

0021-9193

DOI

10.1128/jb.105.1.376-383.1971

Published

1971-01-00

Pages

376-83

Language

English

Region

United States

NLM ID

2985120R

PMCID

PMC248365

Subset

External Links

PubMed source DOI: 10.1128/jb.105.1.376-383.1971 PMC Full Text View journal details

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Purification of intact flagella from Escherichia coli and Bacillus subtilis.

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