Biopolymers and cell. 2006; 22 (6): 425 - 432

Isolation and purification isoaccepting tRNA₁^Ser and tRNA₂^Ser from Thermus thermophilus

I.A. Krikliviy, O.P. Kovalenko, O.Y. Gudzera, A.D. Yaremchuk, M.A. Tukalo

Serine aminoacylation system is unique as it is the only system where synthetase of class II recognizes tRNA with a long variable stem (type II tRNA) which is of great interest to study the mechanisms of tRNA recognition by corresponding aminoacyl-tRNA synthetase. Current paper presents the methodological approaches for obtaining purified isoacceptor tRNA₁^Ser and tRNA₂^Ser in milligram quantities from extremal thermophil Thermus thermophilus for crystallography. The developed method of tRNA^Ser purification employs the chromatography on benzoylated diethylaminoethylcellulose and high-performance liquid chromatography on anion-exchange column Spherogel TSK DEAE 5PW, and reverse phase column Ultrapore C8 using HPLC equipment (Beckman).

Key words: tRNA, chromatography, high-performance liquid chromatography, extremal thermophil Thermus thermophilus, benzoylated diethylaminoethylcellulose.