Enzymatic Aminoacylation of tRNA^Arg Using Recombinant Arg-tRNA Synthetase

This chapter describes the preparation of pre-charged Arg-tRNA that can be used in arginylation reaction. While in a typical arginylation reaction arginyl-tRNA synthetase (RARS) is normally included as a component of the reaction and continually charges tRNA during arginylation, it is sometimes necessary to separate the charging and the arginylation step, in order to perform each reaction under controlled conditions, e.g., for measuring the kinetics or determining the effect of different compounds and chemicals on the reaction. In such cases, tRNA^Arg can be pre-charged with Arg and purified away from the RARS enzyme prior to arginylation.