A method for coupled transcription and aminoacylation of cysteinyl-tRNA.
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.
Pavel
Ioana
I
Department 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.
Belcher
Angela
A
Browning
Karen S
KS
D049148:Aminoacylation; D001710:Biotin; D004926:Escherichia coli / Q000235:genetics*; D014176:Protein Biosynthesis; D012329:RNA, Bacterial / Q000235:genetics*; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry* / Q000235:genetics*; D014158:Transcription, Genetic
D012329:RNA, Bacterial; D012346:RNA, Transfer, Amino Acyl; D001710:Biotin