Construction of FLAG and histidine tagging vectors for Schizosaccharomycespombe

Schizosaccharomyces pombe is becoming an increasingly popular model system for investigating important cellular processes. To facilitate detection, pu rification and functional studies of St. pombe gene products, Ne constructe d two tagging expression vectors for use in St. pombe. These vectors allow proteins to be expressed ectopically as fusion proteins with a FLAG epitope and six histidine residue tags attached to their N-terminus or C-terminus. The function and applicability of these vectors were examined and the resu lts are shown using the N-terminal tagging vector encoding Sfc6p, a subunit of the St. pombe RNA polymerase m general transcription factor, TFIIIC. Co pyright (C) 2001 John Wiley & Sons, Ltd.