Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Amino Acids / analysis
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Animals
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Base Sequence
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Carcinoma, Hepatocellular / genetics
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Chromatography, Affinity
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Chromatography, Ion Exchange
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DNA
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Electrophoresis, Polyacrylamide Gel
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Humans
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Immunodiffusion
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Liver Neoplasms / genetics
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Molecular Sequence Data
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Plasmids / genetics
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Protein Sorting Signals / genetics
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Radioimmunoassay
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Rats
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Saccharomyces cerevisiae / genetics*
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Transfection
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alpha-Fetoproteins / biosynthesis*
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alpha-Fetoproteins / genetics
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alpha-Fetoproteins / isolation & purification
Substances
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Amino Acids
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Protein Sorting Signals
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Recombinant Proteins
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alpha-Fetoproteins
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DNA