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En enkel fluorescens-baserte Reporter analysen identifisere cellulære komponenter kreves for Risin gift en kjede (RTA) handel med gjær
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Journal JoVE Biologie
A Simple Fluorescence-based Reporter Assay to Identify Cellular Components Required for Ricin Toxin A Chain (RTA) Trafficking in Yeast

En enkel fluorescens-baserte Reporter analysen identifisere cellulære komponenter kreves for Risin gift en kjede (RTA) handel med gjær

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

Chapitres

  • 00:05Titre
  • 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

Traduction automatique

Traduction automatique

I manuskriptet beskriver vi bruk av en gjær-baserte fluorescens reporter analysen identifisere cellulære komponenter involvert i smugling og drepe prosesser av den cytotoksiske en undergruppe til anlegget gift Risin (RTA).

Tags

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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