Single strand conformational polymorphism evaluation for the identification of TH3R alleles of the circumsporozoite protein gene of Plasmodium falciparum.
Highly polymorphic regions of the circumsporozoite protein (CSP) of Plasmodium falciparum are associated with cellular immune responses. One of these regions, the TH3R polymorfic region of the csp gene codes for known T-cell epitopes. The present study tested the use of SSCP to determine sequence variations of the TH3R regions of four clones of P. falciparum (3D7, HB3, Dd2 and K1) which are known to have different TH3R regions. Single-strand conformation polymorphism (SSCP) technique was performed on amplified products labelled with fluorescent primer (both strands) and electrophoresed in an automated sequencer. Various gel compositions and electrophoresis conditions were tested. Even if some electrophoretogram differences were observed between clones, they could not distinguish between the alleles.
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Centro de Malária e Outras Doenças Tropicais, UEI Malária, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Rua da Junqueira 96, 1349-008 Lisbon, Portugal. hsilveira@ihmt.unl.pt
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D000483:Alleles; D000818:Animals; D000940:Antigenic Variation / Q000235:genetics; D000953:Antigens, Protozoan / Q000235:genetics* / Q000276:immunology; D016054:DNA, Protozoan / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000235:genetics* / Q000276:immunology; D010963:Plasmodium falciparum / Q000235:genetics* / Q000276:immunology; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D015800:Protozoan Proteins / Q000235:genetics* / Q000276:immunology; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid
D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D000939:Epitopes; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan