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A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies
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JoVE Journal Bioquímica
A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies

A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies

DOI:
10.3791/56080-v

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10:47 min

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July 24, 2017

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, ,
1Department of Neurology,University of Southern California, 2Section of Cell and Developmental Biology,University of California San Diego

Capítulos

  • 00:05Título
  • 00:40Preparation of Competent Cells
  • 02:40Co-transformation of Cells by Heat Shock
  • 04:39Transferring Cells onto New Plates
  • 05:25Preparing Culture for Measuring β-galactosidase Activity
  • 08:14Measuring β-galactosidase Activity
  • 09:07Results: Y1.5H Assay Detects Heteromeric Protein-DNA Interactions
  • 09:50Conclusion

Summary

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The modified yeast one-hybrid assay described here is an extension of the classical yeast one-hybrid (Y1H) assay to study and validate the heteromeric protein complex-DNA interaction in a heterologous system for any functional genomics study.

Tags

Yeast-one HybridHeteromeric Protein ComplexDNA InteractionFunctional GenomicsProtein-DNA InteractionsCo-transformationTE Lithium AcetateYPDA MediumCompetent CellsNormalization Control

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