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Essential Metal Uptake in Gram-negative Bacteria: X-ray Fluorescence, Radioisotopes, and Cell Fractionation
JoVE Journal
Química
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JoVE Journal Química
Essential Metal Uptake in Gram-negative Bacteria: X-ray Fluorescence, Radioisotopes, and Cell Fractionation

Essential Metal Uptake in Gram-negative Bacteria: X-ray Fluorescence, Radioisotopes, and Cell Fractionation

DOI:
10.3791/57169-v

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10:34 min

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February 01, 2018

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, , , , ,
1Graduate Biomedical Sciences Microbiology Theme,University of Alabama at Birmingham, 2Department of Radiology,University of Alabama at Birmingham, 3Office of the Provost,University of Alabama at Birmingham, 4Department of Pharmacology and Toxicology,University of Alabama at Birmingham

Capítulos

  • 00:04Título
  • 01:09Cell Fractionation and Protein Crystallization
  • 03:52X-ray Fluorescence (Using GM/CA Beamline Software)
  • 04:44MAD Scan for Metal Energy Absorption Peak (Using GM/CA Beamline Software)
  • 05:45Analytical Protocol for Radioactive Metal Uptake Assay
  • 07:24Results: Extraction of Bacterial Periplasm from Cytoplasm in Gram-negative Bacteria
  • 09:14Conclusion

Summary

Tadução automática

Tadução automática

A protocol for the extraction of a periplasmic transition metal chaperone in the context of its native binding partners, and biophysical characterization of its substrate contents by X-ray fluorescence and radiometal uptake is presented.

Tags

Keywords: Metal UptakeGram-negative BacteriaX-ray FluorescenceRadioisotopesCell FractionationABC Transport SystemsSubstrate BindingPeriplasmOsmotic ShockSpheroplastsProtein Extraction

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