Isolation and purification isoaccepting tRNA1Ser and tRNA2Ser from Thermus thermophilus

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

doi:10.7124/bc.000748

Biopolym. Cell. 2006; 22(6):425-432.

http://dx.doi.org/10.7124/bc.000748

Structure and Function of Biopolymers

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

1Krikliviy I. A., 1Kovalenko O. P., 1Gudzera O. Y., 1Yaremchuk A. D., 1Tukalo M. A.

Institute of Molecular Biology and Genetics, NAS of Ukraine
150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

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 T. thermophilus for crystallography. The developed method of tRNA purification employs the chromatography on benzoylated diethylaminoethylcellulose and high-performance liquid chromatography on anionexchange column Spherogel TSK DEAE 5PW, and reverse phase column Ultrapore C8 using HPLC equipment (Beckman).

Keywords: tRNA, chromatography, high-performance liquid chro matography, extremal thermophil Thermus thermophilus, benzoyla ted diethylaminoethylcellulose.

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