Expression of human alpha-fetoprotein in yeast.
Human alpha-fetoprotein (AFP) was expressed in Saccharomyces cerevisiae, with a plasmid containing the cDNA sequence for human AFP fused with the rat AFP signal peptide. The recombinant AFP was purified from the yeast lysate by DEAE-cellulose and immunoaffinity chromatography. The amino acid composition and the molecular weight of the recombinant AFP were similar to those of hepatoma AFP. N-terminal amino acids sequence analysis indicated that the signal peptide had been processed. The recombinant and hepatoma AFP reacted identically in Ouchterlony immunodiffusion and radioimmunoassay tests. These observations indicated that the yeast recombinant protein had the properties of native AFP.
Department of Biochemistry, Hokkaido University School of Medicine, Japan.
R
R
Yamamoto
T
T
Sakamoto
S
S
Nishi
M
M
Sakai
T
T
Morinaga
T
T
Tamaoki
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000235:genetics; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D005779:Immunodiffusion; D008113:Liver Neoplasms / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D011863:Radioimmunoassay; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014162:Transfection; D000509:alpha-Fetoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D000596:Amino Acids; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D000509:alpha-Fetoproteins; D004247:DNA