Cloning and Expression of Recombinant Glutathione Peroxidase in Fasciola Gigantica

12 Pages Posted: 10 Oct 2018

Date Written: July 1, 2017

Abstract

Glutathione peroxidase (GPx) is one of the most essential antioxidant enzymes in trematode parasites that lacks catalase. GPx has been already characterized in the trematodes such as Schistoma mansoni, Clonorchis sinensis, and Paragonimus westermani. However, there is no reported about GPx in Fasciola spp. This study amplified GPx cDNA of Fasciola gigantica (FgGPx) from cDNA library. Its sequence contained an in-frame TGA codon for selenocysteine (Sec) which is a stop codon in prokaryote. The Sec (TGA) codon was mutated into a sense codon for cysteine (TGC) using site-directed mutagenesis. The recombinant modified FgGPx (rM_FgGPx) protein was expressed as intracellular protein in Escherichia coli. Polyclonal anti-rM_FgGPx was produced and immunoblotting analysis demonstrated that the anti-rM_FgGPx antibodies did not cross-react with crude antigens from other trematodes including S. mansoni, Opisthorchis viverrini and Paramphistomum spp. The result of ELISA showed that test group had a significant increase of total immunoglobulin and IgG2a when compared with control group. Moreover, there was a significant increase of total immunoglobulin in several F. gigantica infected animal including mice, rabbits and cows, but no cross reaction between O. viverrini infected hamsters sera and rM_FgGPx. Therefore, rM_FgGPx could be used as an immunodiagnostic tool for fascioliasis.

Keywords: Fasciola Gigantica, Glutathione Peroxidase, Mutagenesis, Vaccine Candidate, Immunodiagnostic Tools

Suggested Citation

Abdullohfakeeyah, Muntana and Grams, Hans Rudi and Chunchob, Supatra and Grams, Suksiri Vichasri, Cloning and Expression of Recombinant Glutathione Peroxidase in Fasciola Gigantica (July 1, 2017). PSAKU International Journal of Interdisciplinary Research, Vol. 6, No. 2, Available at SSRN: https://ssrn.com/abstract=3261378

Muntana Abdullohfakeeyah (Contact Author)

Mahidol University ( email )

69 Vipawadee Rangsit Road
Phayatai, Bangkok, Nakhonpathom 10400
Thailand

Hans Rudi Grams

Thammasat University ( email )

Bangkok, 10200
Thailand

Supatra Chunchob

Mahidol University ( email )

69 Vipawadee Rangsit Road
Phayatai, Bangkok, Nakhonpathom 10400
Thailand

Suksiri Vichasri Grams

Mahidol University ( email )

69 Vipawadee Rangsit Road
Phayatai, Bangkok, Nakhonpathom 10400
Thailand

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