title: Using msa-2b as a molecular marker for genotyping Mexican isolates of Babesia bovis.
abstract: Variable merozoite surface antigens of Babesia bovis are exposed glycoproteins having a role in erythrocyte invasion. Members of this gene family include msa-1 and msa-2 (msa-2c, msa-2a(1), msa-2a(2) and msa-2b). To determine the sequence variation among B. bovis Mexican isolates using msa-2b as a genetic marker, PCR amplicons corresponding to msa-2b were cloned and plasmids carrying the corresponding inserts were purified and sequenced. Comparative analysis of nucleotide and deduced amino acid sequences revealed distinct degrees of variability and identity among the coding gene sequences obtained from 16 geographically different Mexican B. bovis isolates and a reference strain. Clustal-W multiple alignments of the MSA-2b deduced amino acid sequences performed with the 17 B. bovis Mexican isolates, revealed the identification of three genotypes with a distinct set each of amino acid residues present at the variable region: Genotype I represented by the MO7 strain (in vitro culture-derived from the Mexico isolate) as well as RAD, Chiapas-1, Tabasco and Veracruz-3 isolates; Genotype II, represented by the Jalisco, Mexico and Veracruz-2 isolates; and Genotype III comprising the sequences from most of the isolates studied, Tamaulipas-1, Chiapas-2, Guerrero-1, Nayarit, Quintana Roo, Nuevo Leon, Tamaulipas-2, Yucatan and Guerrero-2. Moreover, these three genotypes could be discriminated against each other by using a PCR-RFLP approach. The results suggest that occurrence of indels within the variable region of msa-2b sequences can be useful markers for identifying a particular genotype present in field populations of B. bovis isolated from infected cattle in Mexico.
authors.affiliation: Posgrado en Ciencias Genomicas, Universidad Autonoma de la Ciudad de Mexico, Mexico DF, Mexico.
authors.forename: Alma D
authors.initials: AD
authors.lastname: Genis
authors.forename: Jocelin
authors.initials: J
authors.lastname: Perez
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Mosqueda
authors.forename: Antonio
authors.initials: A
authors.lastname: Alvarez
authors.forename: Minerva
authors.initials: M
authors.lastname: Camacho
authors.forename: Maria de Lourdes
authors.initials: Mde L
authors.lastname: Muñoz
authors.forename: Carmen
authors.initials: C
authors.lastname: Rojas
authors.forename: Julio V
authors.initials: JV
authors.lastname: Figueroa
mesh_terms: D000595:Amino Acid Sequence; D054458:Amplified Fragment Length Polymorphism Analysis; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics*; D016793:Babesia bovis / Q000235:genetics* / Q000302:isolation & purification; D001404:Babesiosis / Q000469:parasitology*; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000469:parasitology*; D016054:DNA, Protozoan / Q000032:analysis / Q000235:genetics; D005819:Genetic Markers; D014644:Genetic Variation; D018503:Genome, Protozoan; D008565:Membrane Proteins / Q000235:genetics*; D008800:Mexico; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D005819:Genetic Markers; C476548:MSA-2 protein, Babesia bovis; D008565:Membrane Proteins; D015800:Protozoan Proteins