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A Comparative Approach to Characterize the Landscape of Host-Pathogen Protein-Protein Interactions
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Imunologia e Infecção
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JoVE Journal Imunologia e Infecção
A Comparative Approach to Characterize the Landscape of Host-Pathogen Protein-Protein Interactions

A Comparative Approach to Characterize the Landscape of Host-Pathogen Protein-Protein Interactions

DOI:
10.3791/50404-v

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13:56 min

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July 18, 2013

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, , , ,
1Unité de Génétique, Papillomavirus et Cancer Humain (GPCH),Institut Pasteur , 2Cellule Pasteur,Université Sorbonne Paris Cité, 3Center for Cancer Systems Biology (CCSB), Harvard Medical School, Department of Cancer Biology,Dana Farber Cancer Institute

Capítulos

  • 00:05Título
  • 02:02Yeast Two-hybrid Screenings
  • 05:21Screening of HIS3 Positive Clones by PCR and Sequencing
  • 07:00Cell Transfection
  • 08:26Cell Lysis and Gaussia Luciferase Dosage
  • 09:23Measurement of Firefly Activity and Normalized Luminescence Ratio (NLR) Calculations
  • 10:29Results: Characterization of Virus-host Protein-protein Interactions for the E2, E6 and E7 Early Proteins of Human Papillomaviruses
  • 13:05Conclusion

Summary

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This article focuses on the identification of high-confident interaction datasets between host and pathogen proteins using a combination of two orthogonal methods: yeast two-hybrid followed by a high-throughput interaction assay in mammalian cells called HT-GPCA.

Tags

Protein-protein InteractionsHost-pathogen InteractionsComparative InteractomicsYeast Two-hybridGaussia Luciferase Complementation AssayHigh-throughput ScreeningPathogen Strain VariantsProtein Interaction Networks

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