Abstract
Chemically synthesized genes encodingEscherichia coli tRNA Leu1 and tRNA Leu2 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/A260, respectively: the content of tRNA Leu1 was 50% of total tRNA from MT-Leu1, while that of tRNA Leu2 was 30% of total tRNA from MT-Leu2. Both tRNALeus from their rotal tRNs were fractionated to 1 600 pmol/A260 after DEAE-Sepharose and BD-cellulose column chromatography. The accurate kinetic constants of aminoacylation of the two isoacceptors of tRNALeu catalyzed by leucyl-tRNA synthetase were determined.
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Project supported by the National Natural Science Foundation of China (Grant No. 39570164).
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Li, Y., Wang, E. & Wang, Y. Overproduction and purification ofEscherichia coli tRNALeu . Sci. China Ser. C.-Life Sci. 41, 225–231 (1998). https://doi.org/10.1007/BF02895095
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DOI: https://doi.org/10.1007/BF02895095