Cloning and expression of paraflagellar ROD protein 2 (PFR2) gene of Trypanosoma evansi
2011
Maharana, B.R. | Rao, J.R. | Tewari, A.K. | Siṅgha, Harakīrata
Since Trypanosomes can effectively evade the host immune response by displaying an array of variable surface glycoproteins, attempts for developing a protective immunogen has not been met with success. Owing to its strategic location and invariable nature PFR1 and PFR2, the two important constituent proteins of kinetoplastid flagellum, are considered as impressive candidates for vaccine development against Trypanosomes. In the present study PFR2 gene was cloned in pDRIVE vector. The Histidine tagged recombinant PFR2 was expressed in BL21 cells of E. coli using pET32a vector and subsequently purified using nickel affinity chromatography. The purity of recombinant protein was confirmed by gel electrophoresis. Further confirmation of the recombinant protein based on the immunoreactivity against the tagged Histidine residues was done by western blotting. The nucleotide sequence showed 99.9%, 82.4%, 75.3% and 74.8% sequence homology with the published sequence of Trypanosoma brucei, T. cruzi, Leishmania infantum and Crithidia fasciculata, respectively. The conserved nature of various PFR2 genes present in kinetoplastids is suggestive of its selection as a vaccine candidate against multiple Trypanosoma species.
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