Plasmodium chabaudi: expression of active recombinant chabaupain-1 and localization studies in Anopheles sp.
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.
Caldeira
Rubina L
RL
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.
Gonçalves
Lídia M D
LM
Martins
Tiago M
TM
Silveira
Henrique
H
Novo
Carlos
C
Rosário
Virgílio do
Vd
Domingos
Ana
A
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
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases