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A Simple Fluorescence-based Reporter Assay to Identify Cellular Components Required for Ricin Toxin A Chain (RTA) Trafficking in Yeast
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A Simple Fluorescence-based Reporter Assay to Identify Cellular Components Required for Ricin Toxin A Chain (RTA) Trafficking in Yeast

A Simple Fluorescence-based Reporter Assay to Identify Cellular Components Required for Ricin Toxin A Chain (RTA) Trafficking in Yeast

DOI:
10.3791/56588-v

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09:03 min

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December 15, 2017

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,
1Molecular and Cell Biology, Department of Biosciences and Center of Human and Molecular Biology (ZHMB),Saarland University

Chapters

  • 00:05Title
  • 00:46Heterologous Expression of His-tagged RTA in E. coli
  • 02:12Purification of His-tagged RTA via Affinity Chromatography
  • 04:21Yeast Transformation and Cell Wall Removal
  • 05:28GFP Reporter Assay Measurement in 96-well Plates
  • 06:57Results: Assay to Identify Cellular Components Required for RTA Trafficking in Yeast
  • 08:30Conclusion

Summary

Automatic Translation

Automatic Translation

In the manuscript, we describe the use of a yeast-based fluorescence reporter assay to identify cellular components involved in the trafficking and killing processes of the cytotoxic A subunit of the plant toxin ricin (RTA).

Tags

Keywords: Fluorescence-based Reporter AssayRicin Toxin A Chain (RTA)YeastCellular ComponentsTraffickingIntercellular TransportRibosomeCytotoxic SubunitPlasma MembraneCytosomeCell TransformationLB Kanamycin MediumOptical DensityIPTGCentrifugationBinding BufferSonicationAffinity PurificationNickel-based ColumnHIS-tagged RTA

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