title: Overproduction and purification ofEscherichia coli tRNA(Leu).
abstract: Chemically synthesized genes encodingEscherichia coli tRNA (1) (Leu) and tRNA (2) (Leu) were ligated into the plasmid pTrc99B. then transformed intoEscherichia coli MT102, respectively. The positive transformants, named MT-Leu1 and MT-Leu2, were confirmed by DNA sequencing, and the conditions of cultivation for the two transformants were optimized. As a result, leucinc accepting activity of their total tRNA reached 810 and 560 pmol/A(260), respectively: the content of tRNA (1) (Leu) was 50% of total tRNA from MT-Leu1, while that of tRNA (2) (Leu) was 30% of total tRNA from MT-Leu2. Both tRNA(Leu)s from their rotal tRNs were fractionated to 1 600 pmol/A(260) after DEAE-Sepharose and BD-cellulose column chromatography. The accurate kinetic constants of aminoacylation of the two isoacceptors of tRNA(Leu) catalyzed by leucyl-tRNA synthetase were determined.
authors.affiliation: Statc Key Labratory of Molecular Biology, Shanghai Institute of Biochemistry, Chinese Academy of Sciences, Shanghai, China.
authors.forename: L
authors.initials: L
authors.lastname: Yong
authors.forename: W
authors.initials: W
authors.lastname: Enduo
authors.forename: W
authors.initials: W
authors.lastname: Yinglai