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

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

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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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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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