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Expression, Solubilization, and Purification of Eukaryotic Borate Transporters
JoVE Journal
Bioquímica
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JoVE Journal Bioquímica
Expression, Solubilization, and Purification of Eukaryotic Borate Transporters

Expression, Solubilization, and Purification of Eukaryotic Borate Transporters

DOI:
10.3791/59166-v

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08:55 min

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March 07, 2019

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, ,
1Department of Biology,Davidson College

Capítulos

  • 00:04Título
  • 00:28Borate Transporter Overexpression in S. cerevisiae
  • 01:55Yeast Membrane Harvest
  • 03:51Protein Solubilization and Purification
  • 05:56Glutaraldehyde Cross-Linking Assay
  • 07:09Results: Representative Borate Transporter Purification and Analysis
  • 08:18Conclusion

Summary

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Here we present a protocol to express, solubilize, and purify several eukaryotic borate transporters with homology to the SLC4 transporter family using yeast. We also describe a chemical cross-linking assay to assess the purified homomeric proteins for multimeric assembly. These protocols can be adapted for other challenging membrane proteins.

Tags

Keywords: Membrane ProteinsSaccharomyces CerevisiaeEukaryotic Expression SystemBorate TransportersProtein PurificationYeast CultureProtein InductionYeast Membrane IsolationBead BeatingProtein Solubilization

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