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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
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
IsolationTranslating RibosomesPeptidyl-tRNAsFunctional AnalysisStructural AnalysisAntibioticsProtein Synthesis InhibitionNascent Polypeptide SynthesisPeptide Bond Formation InhibitionTranslation Termination InhibitionTranslational ArrestGene Expression RegulationRibosome Function AlterationMRNA PurificationBiotin-labeled MRNA
