Purification of soybean dna dependent rna polymerase i on a column of plasmid phfk 206 covalently attached to agarose

Grossmann, K.; Seitz, H.U.; Friedrich, H.

Zeitschrift fuer Naturforschung Section C Journal of Biosciences 39(6): 652-655


Accession: 006233648

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The plasmid pHFK206 consisted of plasmid pBR 322 and an 1,3 .times. 10-6 D insert, a cloned segment of a soybean rRNA repeating unit with a preferential binding region for soybean RNA polymerase I, was coupled to cyanogen bromide activated agarose (Sepharose 4B) and used for affinity chromatography of RNA polymerases. Elution profiles and sodium dodecylsulfate/polyacrylamide slab gels showed that highly purified RNA polymerase from Escherichia coli MRE 600 bound to pHFK 206-Sepharose as holoenzyme with an apparently full complement of .delta.-subunit. With initially purified soybean RNA polymerase I from chromatin of 2,4-D treated hypocotyls, pHFK 206-Sepharose could be used as an affinity chromatography method for purification of soybean RNA polymerase I in high degrees and economy of time.