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Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins
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Genetik
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JoVE Journal Genetik
Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins

Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins

DOI:
10.3791/59635-v

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11:34 min

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August 09, 2019

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1Department of Molecular, Cellular and Developmental Biology,University of Colorado at Boulder

Bölümler

  • 00:04Başlık
  • 01:11Generation of DNA Random Library Pool
  • 02:14Synthesis of Pool 0 RNA
  • 04:58RNA-Protein Binding Reaction and Separation of Bound RNA
  • 08:35Reverse Transcription, PCR Amplification, and Protein Binding
  • 09:47Results: Enrichment of PTB-binding RNAs and Splice Site Switching Assay
  • 10:50Conclusion

Özet

Otomatik Çeviri

Otomatik Çeviri

Sequence specificity is critical for gene regulation. Regulatory proteins that recognize specific sequences are important for gene regulation. Defining functional binding sites for such proteins is a challenging biological problem. An iterative approach for identification of a binding site for an RNA-binding protein is described here and is applicable to all RNA-binding proteins.

Etiketler

Sequence SpaceBinding SitesRegulatory RNA-binding ProteinsSaturation MutagenesisProtein-binding SiteDisease DiagnosisTherapyRandom LibraryRNA PoolBinding ReactionHigh AffinitySpecific BindersPCRT7 RNA PolymeraseTranscriptionGel PurificationAutoradiography

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