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Expression, Solubilization, and Purification of Eukaryotic Borate Transporters
JoVE Journal
生化学
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JoVE Journal 生化学
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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, ,
1生物学科,Davidson College

章

  • 00:04標題
  • 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

概要

自動翻訳

自動翻訳

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.

タグ

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

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