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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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生物化学
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JoVE 杂志 生物化学
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

2,372 Views

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07:03 min

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December 23, 2022

DOI:

10.3791/64541-v

DOI:
10.3791/64541-v

•

07:03 min
December 23, 2022

•

19 Views
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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

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.

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

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Cite this Article

Bonchuk, A., Zolotarev, N., Balagurov, K., Arkova, O., Georgiev, P. Pulldown Assay Coupled with Co-Expression in Bacteria Cells as a Time-Efficient Tool for Testing Challenging Protein-Protein Interactions. J. Vis. Exp. (190), e64541, doi:10.3791/64541 (2022).

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