General purpose tagging vectors for fission yeast.
We have designed a series of vectors for use in the fission yeast Schizosaccharomyces pombe that allow fusion of any protein of interest to a triple HA epitope or a GST domain. The HA epitope may be placed at the N terminus or the C terminus under three different versions of the nmt1 promoter, to allow varying levels of gene expression. The GST tag may be placed at the N terminus or C terminus under control of a fully active nmt1 promoter. This family of vectors has compatible restriction sites and modular design, so that the protein under study may be exchanged easily between different plasmids. Using the Cdc19p protein as a test case, we have demonstrated that these plasmids can express functional tagged proteins in the fission yeast cell.
Forsburg
S L
SL
Molecular Biology and Virology Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA. forsburg@salk.edu
Sherman
D A
DA
D000595:Amino Acid Sequence; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics; D000939:Epitopes; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000235:genetics; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000235:genetics; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification; D012568:Schizosaccharomyces / Q000235:genetics*
D018797:Cell Cycle Proteins; D000939:Epitopes; D005656:Fungal Proteins; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase