Isolation of tRNA isoacceptors by affinity chromatography on immobilized bacterial elongation factor Tu.
Elongation factor Tu from Escherichia coli or Thermus thermophilus was immobilized on cyanogen bromide-activated Sepharose. Immobilized elongation factor Tu X GDP could be converted to Tu X GTP, which is able to bind aminoacyl-tRNA. When bulk tRNA from E. coli, baker's yeast, or bovine liver was aminoacylated by one amino acid only, the resulting aminoacyl-tRNA could be separated in one step from the rest of the tRNA using an affinity column of immobilized elongation factor Tu X GTP. Specific tRNA isoacceptors can be isolated in amounts sufficient for gel electrophorectic analysis, sequence determinations, and hybridization experiments.
D000596:Amino Acids; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli; D006160:Guanosine Triphosphate / Q000032:analysis; D010444:Peptide Elongation Factor Tu; D010445:Peptide Elongation Factors; D012343:RNA, Transfer / Q000302:isolation & purification*; D013824:Thermus
D000596:Amino Acids; D010445:Peptide Elongation Factors; D006160:Guanosine Triphosphate; D012343:RNA, Transfer; D010444:Peptide Elongation Factor Tu