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Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists
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Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists

Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists

DOI:
10.3791/51085-v

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10:51 min

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November 15, 2013

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, , ,
1Department of Genetics,Albert Einstein College of Medicine , 2Shanghai Key Laboratory of Complex Prescription and MOE Key Laboratory for Standardization of Chinese Medicines, Institute of Chinese Materia Medica,Shanghai University of Traditional Chinese Medicine

Chapters

  • 00:05Title
  • 02:47Construction of PXR and SRC-1 Fusions in Yeast Vectors
  • 03:49Yeast Two-hybrid Assays
  • 05:26X-gal Filter Assay
  • 06:32Liquid β-gal Assay
  • 07:49Results: X-gal Lift and β-gal Liquid Assay Interpretation
  • 09:06Conclusion

Summary

Automatic Translation

Automatic Translation

Ketoconazole binds to and antagonizes Pregnane X Receptor (PXR) activation. Yeast high throughput screens of PXR mutants define a unique region for ketoconazole binding. This yeast-based genetic method discovers novel nuclear receptor interactions with ligands that associate with surface binding sites.

Tags

Keywords: Reverse Yeast Two-hybrid SystemNuclear ReceptorLigandsAntagonistsPregnane X Receptor (PXR)KetoconazoleCoactivator SRC-1Genetic AssayAntagonist Binding Sites

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