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Extraction and Visualization of Protein Aggregates after Treatment of Escherichia coli with a Proteotoxic Stressor
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JoVE Journal Biochimica
Extraction and Visualization of Protein Aggregates after Treatment of Escherichia coli with a Proteotoxic Stressor

Extraction and Visualization of Protein Aggregates after Treatment of Escherichia coli with a Proteotoxic Stressor

DOI:
10.3791/62628-v

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07:59 min

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June 29, 2021

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, , ,
1School of Biological Sciences,Illinois State University

Capitoli

  • 00:05Introduction
  • 00:53Stress Treatment of E. coli Strains MG1655 and CFT073
  • 02:12Collecting Bacterial Cell Samples
  • 02:43Extracting the Insoluble Protein Aggregates
  • 04:19Soluble Protein Sample Preparation
  • 05:30Separation and Visualization of Extracted Protein Aggregates Using SDS-PAGE
  • 06:49Results: Visualization of Protein Aggregates After Treatment of Escherichia coli with a Proteotoxic Stressor
  • 07:32Conclusion

Summary

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This protocol describes the extraction and visualization of aggregated and soluble proteins from Escherichia coli after treatment with a proteotoxic antimicrobial. Following this procedure allows a qualitative comparison of protein aggregate formation in vivo in different bacterial strains and/or between treatments.

Tags

Keywords: Protein AggregationProteotoxic StressEscherichia ColiProtein ExtractionVisualizationCell LysisGlass Bead DisruptionMOPS-g MediumOD600Antimicrobial Compound

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