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Online Journal of Veterinary Research©
(Including Medical and Laboratory Research)
Volume 25 (8):522-548, 2021
Expression of Mycobacterium avium PPE34.9 interferon gene in HeLa cell line in mice.
Rajib Deb, PP Goswami, NS Prasad.
Gene Expression Laboratory, Division of Animal Biotechnology, Indian Veterinary Research Institute, Izatnagar, UP 243122, India.
Deb R, Goswami PP, Prasad NS., Expression of Mycobacterium avium PPE34.9 interferon gene in HeLa cell line in mice, Onl J Vet Res., 25 (8):522-548, 2021. Mycobacterium avium in Johne’s disease is caused by cell mediated immune response. Genome sequencing proline–proline–glutamic acid protein (PPE) are candidate genes. We constructed a bi-cistrionic pIR PPE/IFN 34.9 kDa PPE protein with cytokine gene encoding murine gamma Interferon gene (IFNγ) and a mono-cistrionic pIR PPE mammalian vector system pIRES 6.1. These were then transfected to a HeLa cell line for expression by Western blot and Immunofluorescent assays with recombinant sera. We then determined immunoreactivity of both constructs of DNA in mice by DTH response, LTT, NO production assay and ELISA. DTH response was higher in pIR PPE/IFN than pIR PPE group of mice, with similar findings for LTT and NO assay where pIR PPE/IFN showed higher (P < 0.01) values than pIR PPE group. ELISA titer of pIR PPE/IFN was lower (P < 0.05) than PPE only. Our findings suggest CMI response of PPE Map and IFNγ synergize T cell immunity in mice.
Key Words: PPE 34.9, FJ032182, IFNγ, Bi-cistrionic expression , HeLa cell line.