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 resol ution 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 th e tRNAs result in a considerable shift of retention time on this colum n. The method is useful not only for obtaining tRNA preparations as re quired for poly(U) translational studies, but also for producing 20-50 -mg amounts of tRNA for NMR and X-ray analysis. These aminoacylated sp ecies (charged by crude synthetase mixtures) can be purified from the crude tRNA mixtures in a one-step procedure.