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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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Biochemistry
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JoVE Journal Biochemistry
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

Chapters

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

Summary

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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.

Tags

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

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