Paraflagellar rod proteins required for cell motility are unique among the kinetoplastids and their heteropolymers provide the building block of the flagellum. We investigated the existence of the paraflagellar rod protein 2 (PFR2) gene in Trypanosoma evansi by reverse transcription-polymerase chain reaction (RT-PCR) using primers designed based on the open reading frame of the PFR2 gene of Trypanosoma brucei. The PFR2 gene was cloned and the PFR2-encoded protein was expressed in bacteria. The expressed His-tag protein was purified using nickel affinity chromatography and confirmed by gel electrophoresis and Western blotting. The nucleotide sequence of the PFR2 gene of T. evansi showed 100% identity with the sequence of the PFR2 gene of T. brucei and 83.4% and 76.6% similarity with that of Trypanosoma cruzi and Leishmania mexicana, respectively. The conserved domain among various PFR2 genes present in kinetoplastids could be used as a target for the development of vaccines against multiple Trypanosoma species.