Izvorni znanstveni članak

https://doi.org/10.24099/vet.arhiv.0106

Cloning, expression and characterization of Trypanosoma evansi Paraflagellar Rod 2 gene

S. Sivajothi orcid.org/0000-0002-1720-9308 ; Department of Veterinary Parasitology, Sri Venkateswara Veterinary University, Tirupati, India
V. C. Rayulu ; Department of Veterinary Parasitology, Sri Venkateswara Veterinary University, Tirupati, India
P. M. Kondaiah ; Department of Veterinary Parasitology, Sri Venkateswara Veterinary University, Tirupati, India
D. Sreenivasulu ; Department of Veterinary Microbiology, Sri Venkateswara Veterinary University, Tirupati, India
CH. Srilatha ; Department of Veterinary Pathology, Sri Venkateswara Veterinary University, Tirupati, India
D. V. R. Sai Gopal ; Department of Virology, Sri Venkateswara University, Tirupati, India
B. Bhaskar Reddy ; RARS, ANGRAU, Tirupati, India
B. Sudhakara Reddy ; Department of Veterinary Medicine, Sri Venkateswara Veterinary University, Tirupati, India

Sažetak

Paraflagellar rod is the major structural component of the trypanosomatid flagellum and is identified as a complex lattice of filaments which runs parallel to the axoneme throughout most of the flagellar length. The present study was carried out to investigate the existence of the paraflagellar rod (PFR 2) gene in Trypanosoma evansi infecting Indian cattle. Local isolates of T. evansi collected from naturally infected cow were multiplied in Wistar rats. Complementary DNA (cDNA) was synthesized from the RNA of host cell free T. evansi parasites by reverse transcription. The gel purified PCR product (PFR 2 gene of T. evansi) was cloned into the pTZ57R/T vector system. The nucleotide sequence of the PFR 2 gene of the T. evansi S.V.V.U. isolate (Accession No. KT277497) obtained in the present study revealed 100% homology with T. evansi China isolate and 99% homology with T. evansi Izatnagar and Bikaner isolates. The recombinant protein was sub-cloned into pET 32a and expressed in the BL21 (DE3) pLysS expression system. The PFR 2 gene of T. evansi S.V.V.U. isolate was further characterized by determination of its protein profile with SDS-PAGE and western blotting. Indirect ELISA was optimized for detection of the specific antibody titre against the recombinant protein of the PFR 2 gene of T. evansi. In the kinetoplastid species the PFR 2 gene is highly conserved. Therefore the PFR 2 gene was suggested as a vaccine candidate, as well as a diagnostic antigen.

Ključne riječi

Andhra Pradesh, India; Trypanosoma evansi; cattle; paraflagellar rod (PFR) 2 gene; cloning and expression

Hrčak ID:

217711

URI

https://hrcak.srce.hr/217711

Datum izdavanja:

8.3.2019.

Podaci na drugim jezicima: hrvatski

Posjeta: 1.062 *