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JoVE Journal
Biology

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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

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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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