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Updated: Aug 20, 2026

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses
Published on: February 25, 2011
A method for coupled transcription and aminoacylation of cysteinyl-tRNA
Ioana Pavel1, Angela Belcher, Karen S Browning
1Department of Chemistry Biochemistry, Institute for Cell and Molecular Biology, and Center for Nano- and Molecular Science and Technology, University of Texas at Austin, Austin, TX 78712, USA.
Abstract:
A novel method for coupled transcription and aminoacylation of transfer RNA was developed where Escherichia coli cysteine-specific tRNA (tRNA(cys)) was transcribed and aminoacylated in a single reaction. The cys-tRNA(cys) that was synthesized and aminoacylated using this method was functional in in vitro translation. The cys-tRNA(cys) was further modified with biotin (N-iodoacetyl-N-biotinhexylenediamine) to facilitate detection. The biotin-modified cys-tRNAs(cys) was also functional in in vitro translation, allowing the synthesis and detection of biotin-labeled protein.
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