Sequencing characterization and expression analysis of interferon gamma (IFN-γ) in Rusa deer (Rusa timorensis) and Thai cervidae

Authors

  • Sirinart Chaichanathong
  • Worawidh Wajjwalku
  • Manakorn Sukmak
  • Boripat Siriaroonrat
  • Tarasak Nipanunt
  • Nikorn Thongtip

Keywords:

cervidae, gene expression, interferon gamma, real-time PCR, sequence characterization

Abstract

To date, the interferon gamma (IFN-γ) assay has become an important diagnostic tool which is applied alternatively to define infection of intracellular pathogens. In this study, the molecular characteristics of IFN-γ in Rusa deer (Rusa timorensis), Hog deer (Axis porcinus), Sambar deer (Rusa unicolor) and Eld’s deer (Rucervus eldii) were determined. Among the cervid family, the analysis of IFN-γ cDNA sequences indicated 5 polymorphism nucleotide positions. Deduced IFN-γ amino acid compositions among the cervid group were negligibly different (98-100% homology). Preliminary quantification of IFN-γ mRNA expression in the cervidae by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) was demonstrated. qPCR primers were designed and tested for their ability to quantify IFN-γ mRNA with a suggested detection limit at 103 target molecules. Intra- and inter-assay variations of RT-qPCR for IFN-γ were 1.41 and 4.57, respectively. Expression ratio of IFN-γ mRNA in mitogen-stimulated peripheral blood mononuclear cells (PBMCs) relative to non-stimulated cells after normalization against β2-microglobulin (β2M) in the Rusa deer, Eld’s deer and Hog deer were 6.47±3.39, 22.91±7.21 and 17.66±1.30, respectively. This preliminary demonstration may serve as the basic instrument for further development of rapid, simple and reliable immunodiagnostic methods for the study of cervidae in Thailand.

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How to Cite

Chaichanathong, S., Wajjwalku, W., Sukmak, M., Siriaroonrat, B., Nipanunt, T., & Thongtip, N. (2017). Sequencing characterization and expression analysis of interferon gamma (IFN-γ) in Rusa deer (Rusa timorensis) and Thai cervidae. The Thai Journal of Veterinary Medicine, 47(1), 117–122. Retrieved from https://he01.tci-thaijo.org/index.php/tjvm/article/view/81517

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Section

Short Communications