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Use of biotinylated-cysteinyl-tRNA as a non-RI probe in protein synthesis
H Ohtsuka1, T Yokogawa, H Asahara
1Department of Biomolecular Science, Faculty of Engineering, Gifu University, Japan.
Nucleic Acids Symposium Series
|January 1, 1997
Abstract:
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.