Expression and rapid one-step purification of biologically active His-tagged factor C by Ni²⁺affinity column chromatography

Abstract

Factor C is an unusual extracellular protein capable of inducing cytodifferentiation in certain Streptomyces strains. The protein is produced by Streptomyces griseus 45H at such a low amount that the study of its mode of action was hindered by the shortage of purified protein. We report here the expression of C-terminally hexa-His-tagged factor C in Streptomyces lividans and Escherichia coli. Expression in S. lividans is low while in E. coli it is relatively high, yielding about 5--10 mg of biologically fully active protein per liter culture.

Characterization of the c-Myb-responsive Region and Regulation of the Human Type I Collagen α2 Chain Gene by c-Myb

“…The reason for using this antibody was that the available anti-c-Myb antibodies were directed against the COOH terminus of the protein, which is deleted in our recombinant Myb-HIS, and because the histidine tag does not usually participate in the function of recombinant proteins (35)(36)(37)(38)(39)(40). Moreover, the Penta-HIS antibody easily detected the recombinant protein in Western blot analysis (data not shown).…”