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An Assay for Quantifying Protein-RNA Binding in Bacteria
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Genetics
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JoVE Journal Genetics
An Assay for Quantifying Protein-RNA Binding in Bacteria

An Assay for Quantifying Protein-RNA Binding in Bacteria

DOI:
10.3791/59611-v

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07:02 min

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June 12, 2019

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, , , ,
1Department of Biotechnology and Food Engineering,Technion-Israel Institute of Technology, 2Russell Berrie Nanotechnology Institute,Technion-Israel Institute of Technology

Chapters

  • 00:04Title
  • 00:44Design of Binding-site Plasmids
  • 02:12Design and Construction of the RBP Plasmid
  • 03:11Experiment Setup
  • 04:56Results: Quantifying Protein-RNA Binding in Bacteria
  • 06:32Conclusion

Summary

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In this method, we quantify the binding affinity of RNA binding proteins (RBPs) to cognate and non-cognate binding sites using a simple, live, reporter assay in bacterial cells. The assay is based on repression of a reporter gene.

Tags

Protein-RNA BindingRNA Binding ProteinRBP-RNA AffinityIn VivoPlasmid DesignBinding Site CassetteMCherry Reporter GeneRBP SequenceBacterial TransformationGlycerol Stocks

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