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分离翻译含肽- tRNA的核糖体的功能和结构分析

JoVE Journal
Biology

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JoVE Journal Biology

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses

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分离翻译含肽- tRNA的核糖体的功能和结构分析

DOI:

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11:19 min

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February 25, 2011

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Nitin Shirole, Sreeram Balasubramanian, Charles Yanofsky, Luis Cruz-Vera

¹Department of Biological Sciences,University of Alabama Huntsville, ²Department of Biology,Stanford University

Chapters

00:05Title
02:59Preparation of Cell Free Extracts Depleted of RF2
04:28Preparation of a DNA Template
05:18Preparation of Biotin Labeled mRNA
06:19Isolation of Translating Ribosomes Containing a Peptidyl-tRNA
07:56Analysis of the Isolated Ribosomes Containing the Peptidyl-tRNA
09:23Representative Results
10:52Conclusion

Summary

Automatic Translation

含新生肽- tRNA的核糖体的生化分析的一个主要障碍已被其他核糖体的存在，在同一样品的核糖体不参与翻译的特定基因序列分析。我们制定了一个简单的方法，净化，完全包含利益的新生肽酰- tRNA，核糖体。

Tags

Isolation Translating Ribosomes Peptidyl-tRNAs Functional Analysis Structural Analysis Antibiotics Protein Synthesis Inhibition Nascent Polypeptide Synthesis Peptide Bond Formation Inhibition Translation Termination Inhibition Translational Arrest Gene Expression Regulation Ribosome Function Alteration MRNA Purification Biotin-labeled MRNA

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