Isolation of gram quantities of isoleucyl‐tRNA synthetase from an overproducing strain of Escherichia coli and its use for purification of cognate tRNA

FEBS Letters

Yamada, Hisami¹  Mizushima, Shoji¹  Kawakami, Makoto¹  Miyazaki, Masazumi¹ 

[1] Institute of Molecular Biology, Faculty of Science, Nagoya University, Chikusa-ku, Nagoya 464, Japan

关键词: Isoleucyl-tRNA synthetase;    Overproducing strain;    tRNA isolation;    Complex formation;    PAGE;    polyacrylamide gel electrophoresis;    DTT;    dithiothreitol;    kb;    kilobase pairs;   

DOI  :  10.1016/0014-5793(85)80762-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 tRNA_GAU of greater than 90% purity was easily isolated by taking advantage of a specific complex formation of the synthetase with cognate tRNA.

【 授权许可】

Unknown