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Identifizierung der Bindungsproteine kleiner Liganden mit der differentiellen radialen Kapillarwirkung des Ligandenassays (DRaCALA)
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Identifying the Binding Proteins of Small Ligands with the Differential Radial Capillary Action of Ligand Assay (DRaCALA)
DOI:

09:26 min

March 19, 2021

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Chapters

  • 00:05Introduction
  • 00:42Preparation of Whole Cell Lysates
  • 01:43Nickel-Nitrilotriacetic Acid (Ni-NTA) Affinity Purification of Relseq and GppA
  • 02:55Gel Filtration
  • 03:33Synthesis of 32P-Labeled Guanosine Pentaphosphate (pppGpp) and Guanosine Tetraphosphate (ppGpp)
  • 05:08DRaCALA Screening of the Target Proteins of (p)ppGpp
  • 06:37Quantification and Identification of Potential Target Proteins
  • 07:46Results: DRaCALA Plate Screening and Quantification
  • 08:39Conclusion

Summary

Automatic Translation

Der Differential Radial Capillary Action of Ligand Assay (DRaCALA) kann verwendet werden, um kleine ligandenbindende Proteine eines Organismus unter Verwendung einer ORFeome-Bibliothek zu identifizieren.

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