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Overproduction and purification of recombinant Bacillus subtilis RNA polymerase.
- Source :
-
Protein expression and purification [Protein Expr Purif] 2008 May; Vol. 59 (1), pp. 86-93. Date of Electronic Publication: 2008 Jan 24. - Publication Year :
- 2008
-
Abstract
- 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 omega subunit or containing either the omega(1) (YkzG) or omega(2) (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.
Details
- Language :
- English
- ISSN :
- 1096-0279
- Volume :
- 59
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Protein expression and purification
- Publication Type :
- Academic Journal
- Accession number :
- 18289874
- Full Text :
- https://doi.org/10.1016/j.pep.2008.01.006