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Molecular Cloning, Sequence Analysis, and Immune Recognition of Bm95 from Thai Strains of Rhipicephalus (Boophilus) microplus
Author(s) -
Jittapalapong Sathaporn,
Thanasilp Sarawan,
Kengradomkit Chanya,
Sirinarukmit Teeraphol,
Kaewmongkol Gunn,
Stich Roger W.
Publication year - 2008
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1428.059
Subject(s) - rhipicephalus microplus , biology , complementary dna , amplicon , rhipicephalus , pichia pastoris , recombinant dna , virology , cloning (programming) , molecular cloning , sequence analysis , antigen , gene , genetics , polymerase chain reaction , tick , computer science , programming language
The efficacy of existing vaccines against Thai strains of Rhipicephalus (Boophilus) microplus remains to be determined, and these vaccine antigens should be compared to homologues derived from local strains. The purpose of this study was to compare Bm95 from Thai R. microplus to the previously reported sequence from Argentinean ticks. mRNA was isolated from R. microplus midgut samples, and cDNA was amplified with Bm95‐specific primers. The cDNA and protein sequences of Thai Bm95 were 94 and 92% identical, respectively, to Argentinean Bm95. Although the sequence was similar to Argentinean Bm95, there were 45‐amino acid differences among the homologues. Amplicons encoding Bm95 were cloned into pPICZαA and expressed in Pichia pastoris strain KM71H. The recombinant plasmid Bm95 (rBm95) was reactive to bovine hyperimmune serum to Thai R. microplus midgut, indicating that the Thai rBm95 was immunogenic. Further work is needed to compare the efficacies of Thai and Argentinean rBm95 for protection of cattle against infestation with Thai R. microplus.