Biochemistry and Biotechnology
Wiley InterScience Backfile Collection 1832-2000
Process Engineering, Biotechnology, Nutrition Technology
Gram-sized quantities of purified arginine, formylmethionine, glutamic acid, and phenylalanine-2 tRNAs have been prepared from pools of E. coli K-12 MO7 mixed tRNAs by reversed-phase chromatography after preliminary fractionation on DEAE-cellulose. Purified formylmethionine tRNA and partially purified arginine tRNA and glutamie acid tRNA were obtained from large-scale RPC-3 runs (4 × 36 in. column). The arginine tRNA was further purified by rechromatography on RPC-4 columns, and the gluatmic acid tRNA by rechromatography on an RPC-3 column. Two phenylalanine tRNAs were resolved on large-scale (2 × 96 in. column) RPC-3 runs; only the second phenylalanine tRNA reached a satisfactory degree of activity. About 0.88 g of arginine tRNA, 70% activity; 3.32 g of formylmethionine tRNA, 97% activity; 0.80 g of glutamic acid tRNA, 83% activity and O.92 g of phenylalanine-2 tRNA, 78% activity, were produced. The processing steps employed are reliable and reproducible and the procedure is amenable to routine production of these tRNAs.
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