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mRNA Interactome Capture from Plant Protoplasts
JoVE Journal
Bioquímica
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JoVE Journal Bioquímica
mRNA Interactome Capture from Plant Protoplasts
DOI:

12:29 min

July 28, 2017

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Capítulos

  • 00:05Título
  • 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

Summary

Tadução automática

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.

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