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Separation of the Cell Envelope for Gram-negative Bacteria into Inner and Outer Membrane Fractions with Technical Adjustments for Acinetobacter baumannii
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Separation of the Cell Envelope for Gram-negative Bacteria into Inner and Outer Membrane Fractions with Technical Adjustments for Acinetobacter baumannii
DOI:

10:24 min

April 10, 2020

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Chapters

  • 00:04Introduction
  • 00:55Preparation of Bacteria for Membrane Extraction
  • 02:18Dissociation of the Outer Membrane and Plasmolysis
  • 03:56Pressurized Homogenization and Lysis
  • 04:35Total Membrane Fractionation
  • 05:34Separating the Dual Membranes
  • 08:00Results: Purity and Quality of Inner and Outer Membrane Fractions
  • 09:36Conclusion

Summary

Automatic Translation

Gram-negative bacteria produce two spatially segregated membranes. The outer membrane is partitioned from the inner membrane by a periplasm and a peptidoglycan layer. The ability to isolate the dual bilayers of these microbes has been critical for understanding their physiology and pathogenesis.

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