title: An improved strategy for tandem affinity purification-tagging of Schizosaccharomyces pombe genes.
abstract: Tandem affinity purification (TAP) is a method that allows rapid purification of native protein complexes. We developed an improved technique to fuse the fission yeast genes with a TAP tag. Our technique is based on tagging constructs that contain regions homologous to the target gene cloned into vectors carrying a TAP tag. We used this technique to design strategies for TAP-tagging of predicted Schizosaccharomyces pombe genes (http://mendel.imp.ac.at/Pombe_tagging/). To validate the approach, we purified the proteins, which associated with two evolutionarily conserved proteins Swi5 and Sfr1 as well as three protein kinases Ksg1, Orb6 and Sid1.
authors.affiliation: Max F. Perutz Laboratories, University of Vienna, Vienna, Austria.
authors.forename: Lubos
authors.initials: L
authors.lastname: Cipak
authors.forename: Mario
authors.initials: M
authors.lastname: Spirek
authors.forename: Maria
authors.initials: M
authors.lastname: Novatchkova
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Chen
authors.forename: Cornelia
authors.initials: C
authors.lastname: Rumpf
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lugmayr
authors.forename: Karl
authors.initials: K
authors.lastname: Mechtler
authors.forename: Gustav
authors.initials: G
authors.lastname: Ammerer
authors.forename: Edina
authors.initials: E
authors.lastname: Csaszar
authors.forename: Juraj
authors.initials: J
authors.lastname: Gregan
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012568:Schizosaccharomyces / Q000737:chemistry* / Q000235:genetics; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins; D029702:Schizosaccharomyces pombe Proteins