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mRNA Interactome Capture from Plant Protoplasts
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JoVE Journal Biyokimya
mRNA Interactome Capture from Plant Protoplasts

mRNA Interactome Capture from Plant Protoplasts

DOI:
10.3791/56011-v

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12:29 min

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July 28, 2017

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, , ,
1Biyoloji Bölümü,KU Leuven

Bölümler

  • 00:05Başlık
  • 01:15In Vivo mRNA-protein Crosslinking by UV Irradiation
  • 02:38Protoplast Lysis Under Denaturing Conditions
  • 03:59mRNP Pull-down and Purification by Oligo-d(T)25 Beads
  • 08:14mRBP Concentration and Protein Assays
  • 10:04Results: Capture of UV-crosslinked mRNA-bound Proteins by Oligo-d(T)25 Beads
  • 11:53Conclusion

Özet

Otomatik Çeviri

Otomatik Çeviri

Here, we present an interactome capture protocol applied to Arabidopsis thaliana leaf mesophyll protoplasts. This method critically relies on in vivo UV crosslinking and allows for the isolation and identification of plant mRNA-binding proteins from a physiological environment.

Etiketler

MRNA InteractomeMRNA-bound ProteomePlant ProtoplastsPost-transcriptional Gene RegulationUV CrosslinkingArabidopsis ThalianaLeaf Mesophyll ProtoplastsMMG SolutionCentrifugation

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