PMID:2581811

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Citation

Kawakami, M, Miyazaki, M, Yamada, H and Mizushima, S (1985) Isolation of gram quantities of isoleucyl-tRNA synthetase from an overproducing strain of Escherichia coli and its use for purification of cognate tRNA. FEBS Lett. 185:162-4

Abstract

The ileS gene coding for isoleucyl-tRNA synthetase was cloned on a runaway-replication plasmid. From the cells harboring the plasmid, gram quantities of the synthetase were isolated using two column procedures. The synthetase was used for the purification of cognate tRNA. Isoleucine tRNAGAU of greater than 90% purity was easily isolated by taking advantage of a specific complex formation of the synthetase with cognate tRNA.

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Keywords

Amino Acyl-tRNA Synthetases/isolation & purification; Chromatography; Chromatography, Gel; Electrophoresis, Polyacrylamide Gel; Escherichia coli/enzymology; Isoleucine-tRNA Ligase/isolation & purification; Isoleucine-tRNA Ligase/metabolism; Plasmids; RNA, Bacterial/isolation & purification; RNA, Transfer, Amino Acyl/isolation & purification; RNA, Transfer, Amino Acyl/metabolism

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