Overproduction and purification of lysyl-tRNA synthetase encoded by the herC gene of E coli.
Lysyl-tRNA synthetases are synthesized from two distinct genes in E coli, lysS and lysU, but neither gene product has been purified distinctively by using overproducing systems. The lysS gene has been identified by a herC mutation which restores maintenance of the mutant ColE1 replicon. The herC gene product was overproduced by using a tac promoter fusion and purified to homogeneity. The purified HerC protein possesses a lysyl-tRNA synthetase activity as predicted by the sequence identity of herC to lysS. The procedure is useful for rapid mass-scale purification of lysyl-tRNA synthetase.
Nakamura
Y
Y
Department of Tumor Biology, University of Tokyo, Japan.
Kawakami
K
K
D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005798:Genes, Bacterial; D008250:Lysine-tRNA Ligase / Q000096:biosynthesis* / Q000302:isolation & purification; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
D011994:Recombinant Proteins; D008250:Lysine-tRNA Ligase