Biopolymers and cell. 2009; 25 (6): 445 - 450

Cloning, expression and purification of tRNA^Pro from bacteria Enterococcus faecalis

K. S. Boyarshin, I. A. Kriklivyi, A. D. Yaremchuk, M. A. Tukalo

Institute of molecular biology and genetics NAS of Ukraine, 150, Zabolotnogo Str, Kyiv Ukraine, 03680

Aim. To elaborate the method of expression and purification of bacteria Enterococcus faecalis tRNA^Pro transcript. Methods. tRNA, co-expressed in vitro with cis-hydrolytical ribozyme, was purified by high performance liquid chromatography using anion-exchange chromatographic column. Results. A satisfactory yield of high purity preparation was obtained. A transcript of tRNAPro exhibits acceptor activity in aminoacylation reaction. Conclusions. The method developed may be introduced in laboratory practice including the obtaining of other tRNAs.

Keywords: tRNA, transcript, gene expression in vitro, high performance liquid chromatography.