Abstract
Plasmodium cysteine proteases have been shown to be immunogenic and are being used as malaria potential serodiagnostic markers and vaccine targets. Genes encoding two Plasmodium chabaudi cysteine proteases chabaupain-1 (CP-1) and chabaupain-2 (CP-2) were identified and further expressed in Escherichia coli. Solubilisation of recombinant CP-1 and CP-2 was achieved by decreasing the temperature of induction. Anopheles gambiae tissues infected with Plasmodium were analyzed by Western blotting using the anti-CP-1 antibody showing that CP-1 is only present in the A. gambiae midguts being absent from other infected mosquito biological material. Anti-CP-1 anti-serum recognized a 30 kDa band in P. chabaudi, Plasmodium berghei and Plasmodium yoelii lysates but does not recognize the recombinant CP-2 extracts suggesting high antibody specificity.
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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Animals
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Anopheles / parasitology*
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Antibodies, Monoclonal / biosynthesis
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Antibodies, Monoclonal / immunology
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Blotting, Western
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Cysteine Endopeptidases / analysis*
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Cysteine Endopeptidases / chemistry
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Cysteine Endopeptidases / genetics
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Cysteine Endopeptidases / immunology
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Electrophoresis, Polyacrylamide Gel
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Enzyme-Linked Immunosorbent Assay
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Female
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Gene Expression Regulation, Enzymologic
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Mice
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Mice, Inbred BALB C
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Molecular Sequence Data
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Phylogeny
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Plasmodium chabaudi / enzymology*
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Plasmodium chabaudi / genetics
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Plasmodium chabaudi / immunology
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Protein Folding
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Recombinant Proteins / analysis
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / immunology
Substances
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Antibodies, Monoclonal
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Recombinant Proteins
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Cysteine Endopeptidases