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生物化学
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Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription
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生物化学
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JoVE 杂志 生物化学
Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription

Application of Biolayer Interferometry (BLI) for Studying Protein-Protein Interactions in Transcription

DOI:
10.3791/59687-v

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07:18 min

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July 26, 2019

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, ,
1Department of Pharmacology, Robert Wood Johnson Medical School,Rutgers University, 2Graduate Program in Physiology and Integrative Biology, School of Graduate Studies,Rutgers University, 3Department of Plant Biology, School of Environmental and Biological,Rutgers University

Chapters

  • 00:04Title
  • 00:59Biosensor Hydration and Assay Set-Up
  • 02:51Loading of Ligand onto Biosensor and Washing Away Additional Ligand
  • 03:43Association of Analyte to Ligand and Dissociation of Analyte from Ligand
  • 05:41Results: A 28 Amino Acid Middle Region of GrgA Binds σ28 In Vitro
  • 06:45Conclusion

Summary

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Interactions of transcription factors (TFs) with the RNA polymerase are usually studied using pulldown assays. We apply a Biolayer Interferometry (BLI) technology to characterize the interaction of GrgA with the chlamydial RNA polymerase. Compared to pulldown assays, BLI detects real-time association and dissociation, offers higher sensitivity, and is highly quantitative.

Tags

Biolayer InterferometryBLIProtein-protein InteractionsTranscriptionKinetic ParametersBiosensorNickel NTAReal-time Association And DissociationQuantitative AnalysisPor Dialysis

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