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A vector system for genomic FLAG epitope‐tagging in Schizosaccharomyces pombe
Author(s) -
Noguchi Chiaki,
Garabedian Mikael V.,
Malik Marriam,
Noguchi Eishi
Publication year - 2008
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.200800140
Subject(s) - schizosaccharomyces pombe , epitope , biology , schizosaccharomyces , computational biology , genetics , flag (linear algebra) , genome , fusion protein , gene , saccharomyces cerevisiae , antigen , recombinant dna , mathematics , pure mathematics , algebra over a field
The fission yeast Schizosaccharomyces pombe is a popular model organism to study various cellular processes, although research tools available for S. pombe are relatively inadequate. To facilitate genetic and biochemical investigation in S. pombe , we report here a system of vectors for genomic FLAG epitope‐tagging. These vectors enable us to amplify gene‐targeting fragments for integration into specific loci of the S. pombe genome. All vectors in this report were designed to express FLAG epitope‐tagged proteins from their endogenous genomic loci. Vectors for N‐terminal FLAG epitope‐tagging allow us to control protein expression levels using the wild‐type nmt1 promoter, its weaker derivatives, and the urg1 promoter. These vectors are available with various antibiotic markers including kanMX6 , hphMX6 , natMX6 and bleMX6 , and the his3 + marker. Vectors for C‐terminal FLAG epitope‐tagging were designed to express FLAG‐fusion proteins under the control of their native promoters at their own genomic loci, allowing us to characterize protein functions under physiological conditions. These vectors are available with kanMX6 , hphMX6 , nat‐MX6 and bleMX6 markers. The series of vectors described in this report should prove useful for protein studies in fission yeast.