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Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions
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JoVE Journal Biyokimya
Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions

Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions

DOI:
10.3791/64541-v

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07:03 min

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December 23, 2022

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, , , ,
1Department of the Control of Genetic Processes, Institute of Gene Biology,Russian Academy of Sciences, 2Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Institute of Gene Biology,Russian Academy of Sciences

Bölümler

  • 00:04Introduction
  • 00:56Co-Transformation of Escherichia coli
  • 02:18Expression
  • 03:21Pulldown Assay
  • 04:52Results: Bacterial Co-Expression of Differentially Tagged Proteins
  • 06:15Conclusion

Özet

Otomatik Çeviri

Otomatik Çeviri

Here, we describe a method for the bacterial co-expression of differentially tagged proteins using a set of compatible vectors, followed by the conventional pulldown techniques to study protein complexes that cannot assemble in vitro.

Etiketler

Pulldown AssayProtein-protein InteractionsCo-expressionHeterodimersProtein Complex AssemblyExpression OptimizationRapid ScreeningEscherichia ColiBacterial TransformationIPTG InductionCell LysisAffinity Purification

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