%0 Journal Article
%T Overproduction and purification of Escherichia coli tRNALeu<br>
%A LI Yong
%A WANG Enduo
%A WANG Yinglai
%A <br>
%J 中国科学C辑(英文版)
%D 1998
%I Springer
%X 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/A260, 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 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. Project supported by the National Natural Science Foundation of China (Grant No. 39570164).
%K tRNA
%K 1
%K Leu
%K tRNA
%K 2
%K Leu
%K clonlng
%K high-expression
%K purification<br>
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=180CF3A72E750F3261A8A60EDC957784&aid=C098FD8E696B1A99D1C39B16D2EF5F7D&yid=8CAA3A429E3EA654&vid=2001E0D53B7B80EC&iid=38B194292C032A66&sid=4966445AEEBA9556&eid=FA89360EB995A8AD&journal_id=1674-7305&journal_name=ScienceChina.Lifesciences&referenced_num=0&reference_num=13