title: Plasmodium chabaudi: expression of active recombinant chabaupain-1 and localization studies in Anopheles sp.
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.
authors.affiliation: INETI-UTPAM - National Institute of Engineering, Technology and Innovation, Unit of Protein and Monoclonal Antidody Technology, Estrada do Paço do Lumiar, 22, Lisboa 1649-038, Portugal.
authors.forename: Rubina L
authors.initials: RL
authors.lastname: Caldeira
authors.forename: Lídia M D
authors.initials: LM
authors.lastname: Gonçalves
authors.forename: Tiago M
authors.initials: TM
authors.lastname: Martins
authors.forename: Henrique
authors.initials: H
authors.lastname: Silveira
authors.forename: Carlos
authors.initials: C
authors.lastname: Novo
authors.forename: Virgílio do
authors.initials: Vd
authors.lastname: Rosário
authors.forename: Ana
authors.initials: A
authors.lastname: Domingos
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000469:parasitology*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D015153:Blotting, Western; D003546:Cysteine Endopeptidases / Q000032:analysis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010802:Phylogeny; D016788:Plasmodium chabaudi / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D017510:Protein Folding; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases