Isolation of gram quantities of isoleucyl-tRNA synthetase from an overproducing strain of Escherichia coli and its use for purification of cognate tRNA.
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.
Kawakami
M
M
Miyazaki
M
M
Yamada
H
H
Mizushima
S
S
D000604:Amino Acyl-tRNA Synthetases / Q000302:isolation & purification*; D002845:Chromatography; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology*; D007533:Isoleucine-tRNA Ligase / Q000302:isolation & purification* / Q000378:metabolism; D010957:Plasmids; D012329:RNA, Bacterial / Q000302:isolation & purification*; D012346:RNA, Transfer, Amino Acyl / Q000302:isolation & purification* / Q000378:metabolism
D012329:RNA, Bacterial; D012346:RNA, Transfer, Amino Acyl; D000604:Amino Acyl-tRNA Synthetases; D007533:Isoleucine-tRNA Ligase