| Babesiosis caused by Babesia orientalis is one of the most prevalent infections of water buffalo transmitted by Rhipicephalus haemaphysaloides causing a parasitic and hemolytic disease. The organelles proteins localized in apical membrane especially rhoptries neck and microneme protein form a complex called moving junction important during invasion process of parasites belong to apicomplexan group,including Babesia species. The rhoptries neck protein (RON) were firstly found in Toxoplasma gondii and later in Plasmodium falciparum as crucial for parasite invasion and good candidate as potential target for vaccine design because of relative high antigenicity feature. Here by using bioinformatic tools, we identified and characterized the RON2, RON4 and RON5 of Babesia orientalis, the main rhoptryneck proteins of moving junction, actively involved in parasite penetration during host infection which has been shown to form a strong complex interaction in previous reported works on others apicomplexan genera.Additionally the proteins identified were analyzed for the presence of secretory signal peptide, transmembrane domains and scanned for some putative biological function. A truncated fragment coding a 936 bps fragment was cloned in pMD-19T and subcloned into pET32 (a)+ expression vector, expressed in E.coli BL21 strain. Purified recombinant BoRON2 was used to produce polyclonal antibody against BoRON2. Here, we identified the full sequence of gene encoding the rhoptryneck 2 protein that we named BoRON2 which is 4035 bp in full length open reading frame without introns, encoding a polypeptide of 1345 amino acids. Western blot of rBoRON2 probed with buffalo positive serum analysis revealed a band of around 150 kDa in parasite lysates, suggesting an active involvement. These findings most likely are constructive in perspective of ongoing research focused particularly in water buffalo babesiosis prevention and therapeutic; and globally provide new information for genes comparative analysis. |
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