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Protein-tRNA Agarose Gel Retardation Assays for the Analysis of the N6-threonylcarbamoyladenosine TcdA Function
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Biochemistry
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JoVE Journal Biochemistry
Protein-tRNA Agarose Gel Retardation Assays for the Analysis of the N6-threonylcarbamoyladenosine TcdA Function

Protein-tRNA Agarose Gel Retardation Assays for the Analysis of the N6-threonylcarbamoyladenosine TcdA Function

DOI:
10.3791/55638-v

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08:03 min

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June 21, 2017

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, , , ,
1Department of Physical Chemical Biology, Center for Biological Research (CIB-CSIC),Spanish National Research Council (CSIC), 2Department of Immunology,Complutense University School of Medicine, 3Abvance Biotech srl

Chapters

  • 00:05Title
  • 01:13Preparation of the tRNA
  • 02:44Lysis, Extraction, and Purification of tRNA(UUU)
  • 04:05Sample Preparation
  • 05:09Gel Electrophoresis and Staining to Observe tRNA-Protein Interaction
  • 06:33Results: Evaluation of the Interaction Between TcdA and tRNA(UUU)
  • 07:22Conclusion

Summary

Automatic Translation

Automatic Translation

A protocol is presented for the production of tRNA(UUU) and the analysis of tRNA(UUU) in complex with the enzyme TcdA by agarose gel retardation assays.

Tags

Keywords: Protein-tRNA InteractionAgarose Gel Retardation AssayRNA-binding ProteinsTcdA FunctionRNA BiologyProtein-RNA ComplexesE. Coli BL21 DE3PET-23D EC Triple-UIPTG InductionCell LysisAcid Phenol Extraction

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