Abstract
Objective:
To achieve high expression of human renal cell carcinoma-associated antigen G250 in Escherichia coli.
Methods:
The gene fragments encoding the protein obtained by PCR was cloned into prokaryotic expression vector pET32a(+) and expressed in E. coli Rosseta. The immunogenicity of the recombinant protein was evaluated by Western blotting.
Results:
The plasmid pET32a(+)/G250 was constructed and expressed in E. coli Rosseta successfully. Western blot analysis showed that the recombinant protein could be specifically recognized by monoclonal antibody M75.
Conclusion:
Efficient G250 expression is achieved in prokaryotic expression system, which may facilitate further functional study of the protein and its monoclonal antibody preparation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antibodies, Monoclonal / immunology
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Antibody Specificity / immunology
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Antigens, Neoplasm / genetics
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Antigens, Neoplasm / immunology
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Antigens, Neoplasm / metabolism*
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Biomarkers, Tumor / genetics
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Biomarkers, Tumor / immunology
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Biomarkers, Tumor / metabolism
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Blotting, Western
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Carbonic Anhydrase IX
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Carbonic Anhydrases / genetics
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Carbonic Anhydrases / immunology
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Carbonic Anhydrases / metabolism*
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Cloning, Molecular
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Escherichia coli / genetics*
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Gene Expression
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Humans
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
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Recombinant Fusion Proteins / metabolism*
Substances
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Antibodies, Monoclonal
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Antigens, Neoplasm
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Biomarkers, Tumor
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Recombinant Fusion Proteins
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CA9 protein, human
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Carbonic Anhydrase IX
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Carbonic Anhydrases