Comparison of different signal peptides for secretion of heterologous proteins in fission yeast.
In the fission yeast Schizosaccharomyces pombe, there are relatively few signal peptides available and most reports of their activity have not been comparative. Using sequence information from the S. pombe genome database we have identified three putative signal peptides, designated Cpy, Amy and Dpp, and compared their ability to support secretion of green fluorescent protein (GFP). In the comparison we also included the two well-described secretion signals derived from the precursors of, respectively, the Saccharomyces cerevisiae alpha-factor and the S. pombe P-factor. The capability of the tested signal peptides to direct secretion of GFP varied greatly. The alpha-factor signal did not confer secretion to GFP and all the produced GFP was trapped intracellular. In contrast, the Cpy signal peptide supported efficient secretion of GFP with yields approximating 10 mg/L. We also found that the use of an attenuated version of the S. cerevisiae URA3 marker substantially increases vector copy number and expression yield in fission yeast.
Kjaerulff
Søren
S
Pharmexa A/S, Kogle Allé 6, DK-2970, Hørsholm, Denmark. sku@my.molbio.ku.dk
Jensen
Martin Roland
MR
D043402:Cathepsin A / Q000737:chemistry; D005656:Fungal Proteins / Q000235:genetics; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006801:Humans; D010957:Plasmids; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
D005656:Fungal Proteins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D029702:Schizosaccharomyces pombe Proteins; D014409:Tumor Necrosis Factor-alpha; D049452:Green Fluorescent Proteins; D043402:Cathepsin A; C468627:cpy1 protein, S pombe