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Expression and purification of untagged and histidine-tagged folate-dependent tRNA:m5U54 methyltransferase from Bacillus subtilis
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.
https://hal-polytechnique.archives-ouvertes.fr/hal-00805069
Contributor : Denis Roura Connect in order to contact the contributor
Submitted on : Friday, July 9, 2021 - 8:21:05 PM
Last modification on : Friday, January 21, 2022 - 3:26:52 AM
Long-term archiving on: : Sunday, October 10, 2021 - 8:00:51 PM
Contributor : Denis Roura Connect in order to contact the contributor
Submitted on : Friday, July 9, 2021 - 8:21:05 PM
Last modification on : Friday, January 21, 2022 - 3:26:52 AM
Long-term archiving on: : Sunday, October 10, 2021 - 8:00:51 PM
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