title: Pseudo-affinity column chromatography based rapid purification procedure for T7 RNA polymerase.
abstract: Based on the observation that T7 RNAP binds reversibly to the polyaromatic sulphonated triazine dye, cibacron blue, we have developed a rapid purification procedure for T7 RNAP. It employs chromatography of the ammonium sulfate fraction through a blue sepharose column, which has the dye coupled to the solid sepharose support. The enzyme can be eluted by 2M NaCl or 1M NaCl together with 1 mM UTP. These methods are compared with another purification procedure using ion-exchange column chromatography. All of them yield essentially pure T7 RNAP with high specific activity.
authors.affiliation: Biophysics Division, Saha Institute of Nuclear Physics, Calcutta, India.
authors.forename: M
authors.initials: M
authors.lastname: Das
authors.forename: D
authors.initials: D
authors.lastname: Dasgupta
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D002852:Chromatography, Ion Exchange; D004396:Coloring Agents / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D014227:Triazines / Q000378:metabolism; D014764:Viral Proteins
chemical_list: D004396:Coloring Agents; D014227:Triazines; D014764:Viral Proteins; C012842:Cibacron Blue F 3GA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases