Production and secretion of recombinant proteins in Dictyostelium discoideum.
We have expressed useful amounts of three recombinant proteins in a new eukaryotic host/vector system. The cellular slime mold Dictyostelium discoideum efficiently secreted two recombinant products, a soluble form of the normally cell surface associated D. discoideum glycoprotein (PsA) and the heterologous protein glutathione-S-transferase (GST) from Schistosoma japonicum, while the enzyme beta-glucuronidase (GUS) from Escherichia coli was cell associated. Up to 20mg/l of recombinant PsA and 1mg/l of GST were obtained after purification from a standard, peptone based growth medium. The secretion signal peptide was correctly cleaved from the recombinant GST- and PsA-proteins and the expression of recombinant PsA was shown to be stable for at least one hundred generations in the absence of selection.
Dittrich
W
W
School of Biological Sciences, Macquarie University, Sydney, NSW, Australia.
Williams
K L
KL
Slade
M B
MB
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular / Q000379:methods*; D003062:Codon / Q000235:genetics; D017931:DNA Primers; D004023:Dictyostelium; D005822:Genetic Vectors; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D045322:Photosynthetic Reaction Center Complex Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D045331:Photosystem I Protein Complex; D010957:Plasmids; D016133:Polymerase Chain Reaction; D021382:Protein Sorting Signals / Q000096:biosynthesis / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D015183:Restriction Mapping; D012549:Schistosoma japonicum / Q000201:enzymology / Q000235:genetics; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D003062:Codon; D017931:DNA Primers; C073973:PSI-A protein, Synechococcus; D045322:Photosynthetic Reaction Center Complex Proteins; D045331:Photosystem I Protein Complex; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D005982:Glutathione Transferase; D005966:Glucuronidase