FIG. 5.

Aminoacylation of unfractionated tRNA, pure tRNA^Gln, tRNA^Asp, and tRNA^Asn from P. aeruginosa PAO1. The reactions shown in panels A, C, and E were conducted in the presence of unfractionated tRNA and catalyzed by partially purified GlnRS, pure GluRS, and pure AspRS from P. aeruginosa PAO1, respectively. Panel B shows the glutaminylation of pure tRNA^Gln by partially purified GlnRS, and panel F shows the aspartylation of pure tRNA^Asp and tRNA^Asn by pure AspRS. (D) Identification by TLC (see Methods) of the amino acid charged on tRNA by GluRS in the reaction shown in panel C (lane 1) and by GlnRS in the reaction shown in panel A (lane 2). Shown are complete reactions (filled circles) and controls without either tRNA (X) or enzyme (empty circles).