PortEco logo

PMID:19963062

From EcoliWiki
Jump to: navigation, search

Contents

Citation

Vrentas, CE, Gaal, T, Burgess, RR and Gourse, RL (2010) An improved procedure for the purification of the Escherichia coli RNA polymerase omega subunit. Protein Expr. Purif. 71:190-4

Abstract

We report an improved procedure for purification of the omega subunit of Escherichia coli RNA polymerase. In contrast to the original procedure, the revised procedure: (i) allows purification of omega entirely from the soluble fraction, obviating the need for denaturation/renaturation, (ii) results in >99% pure omega in only two chromatographic steps, and (iii) improves the yield of purified omega by at least 5-fold. Reconstitution of E. coli RNAP from omega purified by this procedure, as well as purified sigma and core RNAP lacking omega, produces active holoenzyme in vitro, and co-overexpression of omega from a plasmid containing rpoZ and an additional plasmid encoding the other RNAP core subunits results in production of active core enzyme in vivo.

Links

PubMed PMC2843781 Online version:10.1016/j.pep.2009.11.009

Keywords

DNA-Directed RNA Polymerases/genetics; DNA-Directed RNA Polymerases/isolation & purification; DNA-Directed RNA Polymerases/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Escherichia coli/isolation & purification; Plasmids/genetics; Plasmids/metabolism

Significance

You can help EcoliWiki by summarizing why this paper is useful

Useful Materials and Methods

You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.

Annotations

Gene product Qualifier GO ID GO term name Evidence Code with/from Aspect Notes Status

EcoliWiki Links

Add links to pages that link here (e.g. gene, product, method pages)

See also

References

See Help:References for how to manage references in EcoliWiki.