Results for DNA & Clones ( 997 )
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The pY-SUMOstar vector allows for the intracellular expression of SUMOstar fusion proteins in <em>Saccharomyces cerevisiae</em>. SUMOstar fusion constructs are not processed <em>in vivo</em>, allowing for the same enhancement of protein expression, solubility, and stability afforded by SUMOpro.Protein expression is driven by the <em>S. cerevisiae</em> copper metallothionein (CUP1) promoter, which is both tightly regulated and highly inducible. The plasmid is maintained by the 2-micron element and selected for by the restoration of L-tryptophan biosynthesis in Trp auxotrophs.
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The pY-secSUMOstar vector enables expression and secretion of SUMOstar fusions in Pichia pastoris. Protein expression is driven by the primary alcohol oxidase (AOX1) promoter, which is both tightly regulated and highly inducible. The alpha mating factor (S. cerevisiae) targets the fusion protein to the secretory pathway, while both a FLAG and a His6 tag preceed SUMOstar for convenient purification. Chromosomal integration is accomplished by DNA linearization within the promoter using a variety of convient endonuclease restriction sites, while mult-copy integrants can be obtained through strain selection against increasing concentrations of the antibiotic Zeocin.