Expression and rapid one-step purification of biologically active His-tagged factor C by Ni(2+) affinity column chromatography.
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.
Department of Human Genetics, Medical and Health Science Center, University of Debrecen, Hungary.
Z
Z
Birkó
F
F
Schauwecker
F
F
Pfennig
F
F
Szeszák
S
S
Vitális
U
U
Keller
S
S
Biró
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli; D005798:Genes, Bacterial; D006639:Histidine / Q000378:metabolism; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D013305:Streptomyces griseus / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D014169:Transformation, Bacterial
D000596:Amino Acids; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; C402566:factor C, Streptomyces; D006639:Histidine