Expression and purification of untagged and histidine-tagged folate-dependent tRNA:m5U54 methyltransferase from Bacillus subtilis - Archive ouverte HAL Access content directly
Journal Articles Protein Expression and Purification Year : 2010

Expression and purification of untagged and histidine-tagged folate-dependent tRNA:m5U54 methyltransferase from Bacillus subtilis

(1) , (2) , (3) , (1)
1
2
3

Abstract

Folate-dependent tRNA m5U methyltransferase TrmFO is a flavoprotein that catalyzes the C5-methylation of uridine at position 54 in the T-Psi-C loop of tRNA in several bacteria. Here we report the cloning and optimization of expression in Escherichia coli BL21 (DE3) of untagged, N-terminus, C-terminus (His)6-tagged TrmFO from Bacillus subtilis. Tagged and untagged TrmFO were purified to homogeneity by metal affinity or ion exchange and heparin affinity, respectively, followed by size-exclusion chromatography. The tag did not significantly alter the expression level, flavin content, activity and secondary structure of the protein. Cop. 2010 Elsevier Inc. All rights reserved.
Fichier principal
Vignette du fichier
PREP-2010-TrmFO-author-format.pdf (5.2 Mo) Télécharger le fichier
Origin : Files produced by the author(s)

Dates and versions

hal-00805069 , version 1 (09-07-2021)

Identifiers

Cite

Djemel Hamdane, Stéphane Skouloubris, Hannu Myllykallio, Béatrice Golinelli-Pimpaneau. Expression and purification of untagged and histidine-tagged folate-dependent tRNA:m5U54 methyltransferase from Bacillus subtilis. Protein Expression and Purification, 2010, 73 (1), pp.83-89. ⟨10.1016/j.pep.2010.04.013⟩. ⟨hal-00805069⟩
97 View
62 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More