COMPLETE PURIFICATION OF TRANSFER-RNA, CHARGED OR MODIFIED WITH HYDROPHOBIC GROUPS, BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY ON A C-4 C-18 COLUMN SYSTEM

Phe-tRNA(Phe), N-acetyl-Phe-tRNA(Phe) and Leu-tRNA(Leu-4) (from brewer's yeast and Escherichia coli) were each separated with baseline resolution from the uncharged tRNA species by using a wide-pore C-4 column and inverse salt gradient elution. The alterations at the 3' end of the tRNAs result in a considerable shift of retention time on this column. The method is useful not only for obtaining tRNA preparations as required for poly(U) translational studies, but also for producing 20-50-mg amounts of tRNA for NMR and X-ray analysis. These aminoacylated species (charged by crude synthetase mixtures) can be purified from the crude tRNA mixtures in a one-step procedure.