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An improved procedure for the purification of the Escherichia coli RNA polymerase ω subunit
- Source :
-
Protein Expression & Purification . Jun2010, Vol. 71 Issue 2, p190-194. 5p. - Publication Year :
- 2010
-
Abstract
- Abstract: We report an improved procedure for purification of the ω subunit of Escherichia coli RNA polymerase. In contrast to the original procedure, the revised procedure: (i) allows purification of ω entirely from the soluble fraction, obviating the need for denaturation/renaturation, (ii) results in >99% pure ω in only two chromatographic steps, and (iii) improves the yield of purified ω by at least 5-fold. Reconstitution of E. coli RNAP from ω purified by this procedure, as well as purified sigma and core RNAP lacking ω, produces active holoenzyme in vitro, and co-overexpression of ω from a plasmid containing rpoZ and an additional plasmid encoding the other RNAP core subunits results in production of active core enzyme in vivo. [Copyright &y& Elsevier]
Details
- Language :
- English
- ISSN :
- 10465928
- Volume :
- 71
- Issue :
- 2
- Database :
- Academic Search Index
- Journal :
- Protein Expression & Purification
- Publication Type :
- Academic Journal
- Accession number :
- 48733209
- Full Text :
- https://doi.org/10.1016/j.pep.2009.11.009