Evidence for the existence of paraflagellar rod protein 2 (PFR2) gene in Trypanosoma evansi and its conservation among other kinetoplastid parasites

• Published in: Experimental parasitology 2008-04, Vol.118 (4), p.614-618
• Main Authors: Abdille, Md.H., Li, Shao Yong, Jia, Yonggen, Suo, Xun, Mkoji, G.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; amino acid sequences; Animals; Base Sequence; Biological and medical sciences; Blotting, Western; Cloning, Molecular; Conserved Sequence; conserved sequences; Crithidia fasciculata; flagellum; Fundamental and applied biological sciences. Psychology; Gene Expression; genes; IPTG; isopropyl-beta- d-thiogalactopyranoside; Kinetoplastida; Kinetoplastida - genetics; Leishmania infantum; Leishmania mexicana; Life cycle. Host-agent relationship. Pathogenesis; Mice; Mice, Inbred BALB C; molecular cloning; Molecular Sequence Data; nucleotide sequences; paraflagellar rod protein 2; parasites; PBS containing 0.1% Tween 20; PBST; PFR2; Phylogeny; proteins; Protozoa; Protozoan Proteins - chemistry; Protozoan Proteins - genetics; Protozoan Proteins - isolation & purification; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Restriction Mapping; Reverse Transcriptase Polymerase Chain Reaction; reverse transcription-polymerase chain reaction; RT-PCR; SDS–PAGE; Sequence Alignment; sequence homology; sodium dodecyl sulphate–polyacrylamide gel electrophoresis; Trypanosoma - genetics; Trypanosoma brucei; Trypanosoma cruzi; Trypanosoma evansi
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2007.11.011

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.