Kumar, AM and Vulimiri, SV and Nayak, R (1994) Rapid purification of tRNA(Lys) from rat liver. In: Biochemistry & Molecular Biology International, 33 (6). pp. 1081-1089.
Full text not available from this repository. (Request a copy)Abstract
Fast protein liquid chromatography (FPLC) system using Mono Q (HR 5/5) anion-exchange column chromatography followed by highly cross-linked urea-polyacrylamide gel electrophoresis (urea-PAGE) was used for the purification of lysine-specific tRNA (tRNA(Lys)) from rat liver. Crude tRNA from rat liver was fractionated with a linear gradient of NaCl (0.3-0.8 M) in triethanolamine-HCl buffer, pH 4.5, and the activity of tRNA(Lys) was found to elute between 0.51 and 0.57 M NaCl. Using this concentration range of NaCl, tRNA(Lys) was refractionated on the same column with a shallow gradient, where a single peak of tRNA(Lys) activity was obtained. tRNA(Lys)-rich fractions recovered from the second run were electrophoretically separated on 16% polyacrylamide-7 M urea gel into one major band and three minor bands. The major band showed a specific activity of 997 pmols/A260 U for tRNALys with a 43-fold purification and approximately 17% recovery. The minor bands displayed negligible or no activity for lysine. tRNA(Lys) obtained by this method was found to be homogeneous by competitive aminoacylation. The advantages of FPLC followed by urea-PAGE in the purification of an amino acid-specific tRNA over conventional column chromatography are discussed.
| Item Type: | Journal Article |
|---|---|
| Publication: | Biochemistry & Molecular Biology International |
| Publisher: | Academic press aust |
| Additional Information: | Copyright of this article belongs to Academic press aust. |
| Department/Centre: | Division of Biological Sciences > Microbiology & Cell Biology |
| Date Deposited: | 11 Apr 2011 10:34 |
| Last Modified: | 11 Apr 2011 10:34 |
| URI: | http://eprints.iisc.ac.in/id/eprint/36717 |
Actions (login required)
 |
View Item |
