Rapid purification of His(6)-tagged Bacillus subtilis core RNA polymerase.
Bacillus subtilis core RNA polymerase, containing a His(6)-fusion to the C-terminus of the beta' subunit, was isolated by Ni-NTA, Superdex 200 gel filtration, and Mono Q anion-exchange chromatography. The purified core enzyme was shown to be free of the major sigma factor(A) and the transcription factors NusA and GreA. The purification procedure can be completed within 1 working day, is scalable, and yields highly purified and active core RNA polymerase.
Anthony
L C
LC
McArdle Laboratory for Cancer Research, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Artsimovitch
I
I
Svetlov
V
V
Landick
R
R
Burgess
R R
RR
D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D002845:Chromatography; D002846:Chromatography, Affinity; D012321:DNA-Directed RNA Polymerases / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D015335:Molecular Probes; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic
D015335:Molecular Probes; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; C108570:beta' subunit of RNA polymerase; D012321:DNA-Directed RNA Polymerases