Molecular cloning, characterization and antigenicity ofBabesiasp. BQ1 (Lintan) (Babesiacf.motasi) apical membrane antigen-1 (AMA-1)

Abstract

SUMMARYApical membrane antigen-1 (AMA-1) has been described as a potential vaccine candidate in apicomplexan parasites. Here we characterize theama-1gene. The full-lengthama-1gene ofBabesiasp. BQ1 (Lintan) (BLTAMA-1) is 1785 bp, which contains an open reading frame (ORF) encoding a 65-kDa protein of 594 amino acid residues; by definition, the 5′ UTR precedes the first methionine of the ORF. Phylogenetic analysis based on AMA-1 amino acid sequences clearly separated Piroplasmida from other Apicomplexa parasites. TheBabesiasp. BQ1 (Lintan) AMA-1 sequence is most closely associated with that ofB. ovataandB. bigemina, with high bootstrap value. A recombinant protein encoding a conserved region and containing ectodomains I and II of BLTAMA-1 was constructed. BLTrAMA-1-DI/DII proteins were tested for reactivity with sera from sheep infected byBabesiasp. BQ1 (Lintan). In Western-blot analysis, nativeBabesiasp. BQ1 (Lintan) AMA-1 proteins were recognized by antibodies raised in rabbits against BLTrAMA-1in vitro. The results of this study are discussed in terms of gene characterization, taxonomy and antigenicity.