A method for coupled transcription and aminoacylation of cysteinyl-tRNA

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.