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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.

DOI:10.1007/978-1-0716-2942-0_13
PMID:37010754
Abstract

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.

摘要

相似文献

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本文引用的文献

1
Inosine modifications in human tRNAs are incorporated at the precursor tRNA level.人类转运RNA中的肌苷修饰是在前体转运RNA水平上掺入的。
Nucleic Acids Res. 2015 May 26;43(10):5145-57. doi: 10.1093/nar/gkv277. Epub 2015 Apr 27.
2
An adenosine deaminase that generates inosine at the wobble position of tRNAs.一种在tRNA摆动位置产生肌苷的腺苷脱氨酶。
Science. 1999 Nov 5;286(5442):1146-9. doi: 10.1126/science.286.5442.1146.