Overproduction and purification of recombinant Bacillus subtilis RNA polymerase

Title: Overproduction and purification of recombinant Bacillus subtilis RNA polymerase
Creator: Yang, Xiao; Lewis, Peter J.
Relation: Protein Expression and Purification Vol. 59, Issue 1, p. 86-93
Publisher Link: http://dx.doi.org/10.1016/j.pep.2008.01.006
Publisher: Elsevier
Resource Type: journal article
Date: 2008
Description: We describe a vector-based system for the production of recombinant Bacillus subtilis RNA polymerase. The recombinant enzyme is C-terminally tagged with nine consecutive histidine residues resulting in about 90% pure enzyme in a single nickel-affinity purification step. The vectors permitted production of recombinant enzyme lacking an ω subunit or containing either the ω₁ (YkzG) or ω₂ (YloH) subunits. In transcription time-course assays all of the recombinant enzymes exhibited identical activity to native RNAP. The modular assembly of the artificial RNA polymerase operon permits ready mutation of any subunit and incorporation into the recombinant enzyme, which will enable new functional/structural studies with this enzyme.
Subject: RNA polymerase; Bacillus subtilis; artificial operon; His-tag; transcription
Identifier: http://hdl.handle.net/1959.13/42875
Identifier: uon:5015
Identifier: ISSN:1046-5928
Language: eng
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