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JoVE Journal Biochemistry
Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases
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Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases

Article DOI: 10.3791/62950-v • 06:30 min • August 27th, 2021
August 27th, 2021 •
1, 1
1Department of Molecular and Cellular Biology, College of Biological Science, University of Guelph

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Summary
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Ubiquitination is a critical protein post-translational modification, dysregulation of which has been implicated in numerous human diseases. This protocol details how phage display can be utilized to isolate novel ubiquitin variants that can bind and modulate the activity of E3 ligases that control the specificity, efficiency, and patterns of ubiquitination.

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Phage Display Ubiquitin Variant Modulators E3 Ligases Binding Characteristics Enzyme Modulators Target Proteins Conventional Display Methods Repetitive Action 2YT Tetracycline Broth Mid-log Phase Coating Solution PB Buffer Phage Library Library Diversity Polyethylene Glycol Sodium Chloride Solution Centrifuge
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