In Vivo mRNA-protein Crosslinking by UV Irradiation
2:38
Protoplast Lysis Under Denaturing Conditions
3:59
mRNP Pull-down and Purification by Oligo-d(T)25 Beads
8:14
mRBP Concentration and Protein Assays
10:04
Results: Capture of UV-crosslinked mRNA-bound Proteins by Oligo-d(T)25 Beads
11:53
Conclusion
TRANSCRIÇÃO
The overall goal of this optimized mRNA interactome capture is to isolate and identify an mRNA-bound proteome from plant cells. This method is applied to arabidopsis thaliana leaf mesophil protoplasts, a cell type used as a versatile tool in vario
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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.