Preparation of an Enriched tRNA Fraction for Arginylation by Expression in E. coli.
作者信息
Avcilar-Kucukgoze Irem, Gamper Howard, Hou Ya-Ming, Kashina Anna S
机构信息
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Department of Biochemistry and Molecular Biology, University of Pennsylvania, Philadelphia, PA, USA.
出版信息
Methods Mol Biol. 2023;2620:101-106. doi: 10.1007/978-1-0716-2942-0_13.
The method described here provides a fast and efficient way to obtain an enriched preparation of tRNA of interest, which is also posttranscriptionally modified by the intracellular machinery of the host cells, E. coli. While this preparation also contains a mixture of total E. coli tRNA, the enriched tRNA of interest is obtained in high yields (milligram) and is highly efficient for biochemical assays in vitro. It is routinely used in our lab for arginylation.
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