Phe-tRNAPhe, N-acetyl-Phe-tRNAPhe and Leu-tRNALeu-4 (from brewer's yeast and Escherichia coli) were each separated with baseline resolution from the uncharged tRNA species by using a wide-pore C4 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.
Research Areas and Centers
- Academic Focus: Center for Infection and Inflammation Research (ZIEL)