Construction of FLAG and histidine tagging vectors for Schizosaccharomyces pombe.
Schizosaccharomyces pombe is becoming an increasingly popular model system for investigating important cellular processes. To facilitate detection, purification and functional studies of Sz. pombe gene products, we constructed two tagging expression vectors for use in Sz. 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 results are shown using the N-terminal tagging vector encoding Sfc6p, a subunit of the Sz. pombe RNA polymerase III general transcription factor, TFIIIC.
Laboratory of Molecular Growth Regulation, National Institute of Child Health and Human Development/NIH, Building 6, Room 3403, 6 Center Drive MSC 2753, Bethesda, MD 20892-2753, USA. huangy@exchange.nih.gov
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Y
Huang
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Hamada
J
J
Patel
R J
RJ
Maraia
D000595:Amino Acid Sequence; D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D000939:Epitopes / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D006639:Histidine / Q000235:genetics; D008969:Molecular Sequence Data; D009842:Oligopeptides; D010455:Peptides / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics*; D019474:Transcription Factors, TFIII / Q000737:chemistry; D000098832:Transcription Factor TFIIIC
D000939:Epitopes; D009842:Oligopeptides; D010455:Peptides; D011993:Recombinant Fusion Proteins; D019474:Transcription Factors, TFIII; D000098832:Transcription Factor TFIIIC; D006639:Histidine; C085706:FLAG peptide