Premium
Purification of ten transfer riboiiucleic acids from E. coli K‐12 MO
Author(s) -
Weeren H. O.,
Ryon A. D.,
Kelmers A. D.
Publication year - 1972
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260140407
Subject(s) - valine , chemistry , chromatography , lysine , methionine , aspartic acid , transfer rna , alanine , amino acid , extraction (chemistry) , ethanol , arginine , glutamic acid , biochemistry , rna , gene
A flowsheet has been developed which leads to the purification of 10 transfer ribonucleic acids (tRNAs) from E. coli K‐12 MO. Crude tRNAs were recovered by phenol extraction and two ethanol precipitation steps. The initial separation of the crude tRNA mixture was achieved by RPC‐3 reversed‐phase chromatography. Following rechromatography under other conditions, the following tRNAs were recovered at a purity of 70 to 100%: arginine, aspartic acid, glutamic acid, lysine, formylmethionine‐1, formylmethionine‐3, normal methionine, phcnylalanine‐1, phenylalaninc‐2, and valine. Recoveries for these tRNAs ranged from 15 to 60%. The flowsheet was demonstrated with chromatogaphic runs ranging from 1,300 to 250,000 A 260 units per run. The chromatographic steps were simple and readily reproducible.