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探索序列空间,识别调节RNA结合蛋白的结合位点

JoVE Journal
Genetics

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JoVE Journal Genetics

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

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探索序列空间,识别调节RNA结合蛋白的结合位点

DOI:

10.3791/59635-v

•

11:34 min

•

August 09, 2019

•

Ravinder Singh

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

Chapters

00:04Title
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

Summary

Automatic Translation

序列特异性对基因调控至关重要。识别特定序列的调控蛋白对基因调控非常重要。为这种蛋白质定义功能结合位点是一个具有挑战性的生物学问题。此处描述了一种用于识别RNA结合蛋白结合位点的迭代方法,适用于所有RNA结合蛋白。

Tags

Sequence Space Binding Sites Regulatory RNA-binding Proteins Saturation Mutagenesis Protein-binding Site Disease Diagnosis Therapy Random Library RNA Pool Binding Reaction High Affinity Specific Binders PCR T7 RNA Polymerase Transcription Gel Purification Autoradiography

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