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A rapid microbead method for breaking pathogenic mycobacteria: Application in SDS-PAGE and western blot analysis

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Abstract

A rapid method used to obtain cell-free extracts from pathogenic mycobacteria was successfully applied to five slowly growing species. The method, which simply consisted of vortexing of the bacterial suspensions with 130-μm glass microbeads under predefined conditions, resulted in the breaking of bacterial cells as evidenced by electron microscopy and gave about 1 mg/ml of total proteins. The cell-free extracts so obtained were suitable for SDS-PAGE analysis as well as Western blots. In a pilot test system, this method was applied toMycobacterium avium (M. av),M. paratuberculosis (P.tb), and related species isolated from “wood-pigeon” (M. silvaticum) and Crohn's disease (CD). The procedure was used in the analysis of various mycobacterial proteins by SDS-PAGE and Western blotting, and the findings were compatible with recently published data using criteria based on DNA polymorphism and numerical taxonomy.

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Authors and Affiliations

  1. Tuberculosis and Mycobacteriology Unit Institut Pasteur, 75724, Paris, France

    Nalin Rastogi & Valérie Labrousse

  2. Culture Collection Unit Institut Pasteur, 75724, Paris, France

    Claude Barreau

Authors
  1. Nalin Rastogi
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  2. Valérie Labrousse
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  3. Claude Barreau
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Rastogi, N., Labrousse, V. & Barreau, C. A rapid microbead method for breaking pathogenic mycobacteria: Application in SDS-PAGE and western blot analysis. Current Microbiology 24, 311–317 (1992). https://doi.org/10.1007/BF01571100

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  • DOI: https://doi.org/10.1007/BF01571100

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