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A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates
Bölümler
- 00:04Başlık
- 00:30Generation of a Library of Mutants – Doping DNA Template with Non-wild Type Nucleotides
- 01:20Synthesis of RNA
- 05:24Protein Binding Reaction and Separation of Bound RNA
- 08:05Analysis of Iodine-cleaved Phosphorothioate Products for Detection of Mutant Nucleotide Positions
- 09:07Results: Characterization of Protein Binding Site in RNA
- 10:22Conclusion
Proteins that bind specific RNA sequences play critical roles in gene expression. Detailed characterization of these binding sites is crucial for our understanding of gene regulation. Here, a single-step approach for saturation mutagenesis of protein-binding sites in RNA is described. This approach is relevant for all protein-binding sites in RNA.
