A rapid method for the purification of RNA polymerase holoenzyme from Escherichia coli.
A method is described for the rapid purification of RNA polymerase holoenzyme from small amounts of Escherichia coli cells. Chromatography of a crude extract on a single-stranded DNA agarose column followed by gel filtration chromatography gave 95% pure holoenzyme. The enzyme had kinetic characteristics on T7 DNA identical to those of RNA polymerase purified by other more laborious procedures.
Mehrpouyan
M
M
Department of Biochemistry, College of Medicine, East Tennessee State University, Johnson City 37614.
Champney
W S
WS
D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004269:DNA, Bacterial / Q000502:physiology; D004279:DNA, Viral / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D012367:RNA, Viral / Q000235:genetics; D012685:Sepharose; D013604:T-Phages / Q000235:genetics; D013997:Time Factors; D014158:Transcription, Genetic / Q000235:genetics
D004269:DNA, Bacterial; D004279:DNA, Viral; D012367:RNA, Viral; D012685:Sepharose; D012321:DNA-Directed RNA Polymerases