Molecular Characterization of Oligopeptidase B gene of Trypanosoma evansi Isolated from Camel

Trypanosomosis caused by Trypanosoma evansi is a disease of economic significance of camel, commonly known as Tibersa or Surra. The current treatment regimens based on chemotherapy and chemoprophylaxis for camel Trypanosomiasis are limited, and are not ideal as they are often associated with severe side effects. The development of new trypanocidal drug for trypanosomiasis is very difficult and hopes for the development of conventional vaccines are also limited due to the trypanosome's ability to escape the host immune response through antigenic variation. Trypanosomes change their variant surface glycoprotein coat very quickly which is the main hurdle in vaccine development against this parasite. Recent effort towards the development of a vaccine against T. evansi has identified several promising candidate vaccine antigens, including non-variant proteins like Oligopeptidase B of this parasite. Keeping this in view, the Oligopeptidase B gene of the T. evansi isolates was amplified from the infected blood materials by PCR, the amplicon was cloned into pGEM- T Easy vector and subsequently characterization of the gene through sequencing. After cloning screening of recombinants was done by Restriction Enzyme digestion of plasmid DNA using EcoR1 and found that the release of DNA fragment was around 2092 bp. The sequence analysis revealed that Oligopeptidase B gene of T. evansi from India was having the identity of 100% with T. evansi (Germany isolate), 99.6% with T.brucei (USA isolate) and only 75.7% with T. cruzi (USA isolate). The comparison with Leishmania amazonensis (Brazil isolate), Leishmania donovani (USA isolate), Leishmania infantum (UK isolate) and Leishmania major (USA isolate) a member belonging to Trypanosomatidae family showed only 77.6%, 76.2%, 75.9% and 76.2% homology, respectively.