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United States Department of Agriculture

Agricultural Research Service

Research Project: DETECTION AND TYPING OF FOOD-BORNE PATHOGENS

Location: Molecular Characterization of Foodborne Pathogens

Title: Modification of an acetone-sodium dodecyl sulfate disruption method for cellular protein extraction from neuropathogenic Clostridium botulinum

Author
item BHADURI, SAUMYA

Submitted to: Foodborne Pathogens and Disease
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: September 28, 2011
Publication Date: January 3, 2012
Citation: Bhaduri, S. 2012. Modification of an acetone-sodium dodecyl sulfate disruption method for cellular protein extraction from neuropathogenic Clostridium botulinum. Foodborne Pathogens and Disease. 9:172-174.

Interpretive Summary: Studies of the biological, immunological, and biochemical properties of cellular proteins provide valuable information pertaining to neurotoxigenic pathogens such as Clostridium botulinum. These studies are dependent on the efficient preparation of cell-free extracts which provide sufficient cellular proteins for such investigations. An acetone-sodium dodecyl sulfate method was evaluated for extraction of cellular proteins from C. botulinum prepared for characterization of protein composition. The amount of protein extracted per gram of dry weight and the protein profile as revealed by polyacrylamide gel electrophoresis (PAGE)showed that an extract from C. botulinum for this method was comparable to the extracts made by established methods, namely, sonication or agitation with beads. This study demonstrated the reliability and usefulness of this rapid, safe, and inexpensive procedure to prepare cell-free extract from C. botulinum on biochemical analysis and is also suitable for proteomics analysis by two-dimensional PAGE and liquid chromatography. This method is also suitable for the extraction of radioisotope-labeled proteins from C. botulinum safely, since potential biohazard problems caused by the generation of aerosols and contamination of the apparatus are avoided. This technique could be successfully applied to the extraction of proteins from other highly pathogenic bacteria for similar studies.

Technical Abstract: An acetone-sodium dodecyl sulfate (SDS) disruption method was used for the extraction of cellular proteins from neurotoxigenic Clostridium botulinum. The amount of protein extracted per gram of dry weight and the protein profile as revealed by polyacrylamide gel electrophoresis (PAGE) was comparable to the extracts prepared by established methods, namely, sonication or agitation with beads. This method allows safer handling of C. botulinum by avoiding mechanical disruption, the generation of aerosols, and contamination of the apparatus.

Last Modified: 8/19/2014
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