Pamela Foreman

Cloning vectors for the synthesis of epitope-tagged, truncated and chimeric proteins in Saccharomyces cerevisiae

A series of cloning vectors, designated YCpIF, was constructed to facilitate the conditional synthesis of epitope-tagged, truncated and chimeric proteins in Saccharomyces cerevisiae. These vectors contain a translation start codon upstream from a multiple cloning site (MCS) in each of the three reading frames. Protein synthesis is under the control of the GAL1 promoter, which drives transcription when cells are grown on galactose-containing medium, but not when they are grown on glucose-containing medium. Different versions of the vectors contain four different commonly used selectable markers. In addition, YCpIF15, YCpIF16 and YCpIF17 contain a sequence encoding an epitope from influenza virus hemagglutinin upstream from the MCS. These vectors facilitate the addition of this epitope tag to the N terminus of any protein. The epitope is recognized by a commercially available monoclonal antibody.