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

Article DOI: 10.3791/2498-v • 11:20 min • February 25th, 2011
February 25th, 2011 •
1, 1, 2, 1
1Department of Biological Sciences, University of Alabama Huntsville, 2Department of Biology, Stanford University

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Summary
Automatic Translation

A major impediment to biochemical analyses of ribosomes containing nascent peptidyl-tRNAs has been the presence of other ribosomes in the same samples, ribosomes not involved in the translation of the specific mRNA sequence being analyzed. We developed a simple methodology to purify, exclusively, the ribosomes containing the nascent peptidyl-tRNA of interest.

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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