Pseudo-affinity column chromatography based rapid purification procedure for T7 RNA polymerase.

M Das; D Dasgupta

doi:10.1080/10826069808010146

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Pseudo-affinity column chromatography based rapid purification procedure for T7 RNA polymerase.

M Das·D Dasgupta

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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.

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Chromatography, Affinity Chromatography, Ion Exchange Coloring Agents DNA-Directed RNA Polymerases Electrophoresis, Polyacrylamide Gel Triazines Viral Proteins Journal Article

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M Das, D Dasgupta. 1998. Pseudo-affinity column chromatography based rapid purification procedure for T7 RNA polymerase.. https://doi.org/10.1080/10826069808010146

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