Use of biotinylated-cysteinyl-tRNA as a non-RI probe in protein synthesis.
A convenient method for the preparation of biotinylated aminoacyl-tRNA to use in the non-radioisotopic (non-RI) detection of cell-free translation products was developed. After aminoacylation of E. coli tRNA(Cys) with L-cystein, its sulfhydryl group was modified with N-(6-[Biotinamide]hexyl)-3'-(2'-pyridyl dithio) propionamide or 1-Biotin amido-4-(4'-[maleimidomethyl] cyclohexane-carboxamido) butane. These biotin-labelled cysteinyl-tRNA are expected to function as the non-RI probe for protein synthesis equally to or even better than the biotinylated lysyl-tRNA which is now commercially available.
Ohtsuka
H
H
Department of Biomolecular Science, Faculty of Engineering, Gifu University, Japan.
Yokogawa
T
T
Asahara
H
H
Nishikawa
K
K
D001710:Biotin; D002474:Cell-Free System; D003545:Cysteine / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D008958:Models, Molecular; D014176:Protein Biosynthesis; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl; D012350:RNA, Transfer, Cys / Q000378:metabolism*
D011506:Proteins; D012333:RNA, Messenger; D012346:RNA, Transfer, Amino Acyl; D012350:RNA, Transfer, Cys; D001710:Biotin; D003545:Cysteine