tRNATrp from beef lever has been purified by classical chromatographical methods. Total tRNA, prepared on a large scale (total aminoacid acceptance 1280 pmol/A260 unit) was submitted to chromatography on benzoylated-DEAE cellulose, then DEAE Sephadex. The major species accepting tryptophan issued from the second chromatography was aminoacylated with [14C] tryptophan and chromatographed on benzoylated-DEAE cellulose. The tRNA carrying the radioactive label was eluted in the ethanolic region. After stripping, the resulting tRNATrp has an acceptance of 1800 pmol/A260 unit. No isoacceptors could be demonstrated by chromatography of the pure species on RCP 5 in 6 M urea. The yield in pure tRNATrp was currently in the range of 25 to 30 percent of the total tryptophan acceptance of the starting curde tRNA.
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http://dx.doi.org/10.1016/s0300-9084(76)80114-9 | DOI Listing |
tRNATrp from beef lever has been purified by classical chromatographical methods. Total tRNA, prepared on a large scale (total aminoacid acceptance 1280 pmol/A260 unit) was submitted to chromatography on benzoylated-DEAE cellulose, then DEAE Sephadex. The major species accepting tryptophan issued from the second chromatography was aminoacylated with [14C] tryptophan and chromatographed on benzoylated-DEAE cellulose.
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