A vector system that allows simple generation of mutant Escherichia coli RNA polymerase

Title: A vector system that allows simple generation of mutant Escherichia coli RNA polymerase
Creator: Yang, Xiao; Ma, Cong; Lewis, Peter
Relation: NHMRC.1008014
Relation: Plasmid Vol. 75, p. 37-41
Publisher Link: http://dx.doi.org/10.1016/j.plasmid.2014.06.002
Publisher: Academic Press
Resource Type: journal article
Date: 2014
Description: We describe a dual vector-based system for overproduction of recombinant Escherichia coli RNA polymerase (RNAP). A cleavable deca-histidine tag (His10) was incorporated into the C-terminus of the b0 subunit to facilitate protein purification. Unique restriction sites were introduced into the genes encoding the b and b0 subunits (rpoB and rpoC, respectively), facilitating mutation of functionally significant subunit fragments through insertion of modified PCR fragments into the appropriate vector. RNAP with an R275A substitution in the b0 subunit, which is essential for interaction with transcription initiation factor r, was generated and exhibited reduced activity compared to native recombinant RNAP.
Subject: RNA polymerase; Escherichia coli; dual-expression; transcription
Identifier: http://hdl.handle.net/1959.13/1304578
Identifier: uon:20873
Identifier: ISSN:0147-619X
Language: eng
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