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Expression and characterization of glutathione peroxidase of the liver fluke, Fasciola gigantica

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Abstract

Glutathione peroxidase (GPx) is a key member of the family of antioxidant enzymes in trematode parasites including Fasciola spp. Because of its abundance and central role as an anti-oxidant that helps to protect parasites from damage by free radicals released from the host immune cells, it has both diagnostic as well as vaccine potential against fasciolosis. In this study, we have cloned, characterized, and detected the expression of the GPx protein in Fasciola gigantica (Fg). FgGPx (582 bp) was cloned by polymerase chain reaction (PCR) from complementary DNA (cDNA) from an adult fluke. Its putative peptide has no signal sequence and is composed of 168 amino acids, with a molecular weight of 19.1 kDa, and conserved sequences at NVACKUG, FPCNQFGGQ, and WNF. Phylogenetic analysis showed that GPx is present from protozoa to mammals and FgGPx was closely related to Fasciola hepatica GPx. A recombinant FgGPx (rFgGPx) was expressed in Escherichia coli BL21 (DE3) and used for immunizing mice to obtain polyclonal antibodies (anti-rFgGPx) for immunoblotting and immunolocalization. In immunoblotting analysis, the FgGPx was expressed in all stages of F. gigantica (eggs, metacercariae, newly excysted juveniles (NEJ), 4-week-old juveniles, and adults). This mouse anti-rFgGPx reacted with the native FgGPx at a molecular weight of 19.1 kDa in adult whole body (WB) and tegumental antigens (TA) as detected by immunoblotting. The FgGPx protein was expressed at a high level in the tegument, vitelline glands, and eggs of the parasite. Anti-rFgGPx exhibited no cross-reactivity with the other parasite antigens, including Eurytrema pancreaticum, Cotylophoron cotylophorum, Fischoederius cobboldi, Gastrothylax crumenifer, Paramphistomum cervi, and Setaria labiato papillosa. The possibility of using rFgGPx for immunodiagnosis and/or as a vaccine for fasciolosis in animals of economic importance will be explored in the future.

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Funding

This research was financially supported by The Thailand Research Fund for New Scholar to Dr. Narin Changklungmoa (Grant no. MRG5980048), a grant from Burapha University as approved by the National Research Council of Thailand to Dr. Narin Changklungmoa (Grant no. 210/2561), a grant from Mahidol University as approved by the National Research Council of Thailand to Associate Professor Dr. Kulathida Chaithirayanon, and a Senior Research Scholar grant from Thailand Research Fund to Professor Dr. Prasert Sobhon.

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Authors and Affiliations

  1. Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Road, Saen Sook Sub-district, Mueang District, Chonburi, 20131, Thailand

    Narin Changklungmoa, Werachon Cheukamud, Athit Chaiwichien, Supawadee Osotprasit, Tepparit Samrit, Prasert Sobhon & Pornanan Kueakhai

  2. Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd, Bangkok, 10400, Thailand

    Kulathida Chaithirayanon & Prasert Sobhon

Authors
  1. Narin Changklungmoa
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  2. Kulathida Chaithirayanon
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  3. Werachon Cheukamud
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  4. Athit Chaiwichien
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  5. Supawadee Osotprasit
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  6. Tepparit Samrit
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  7. Prasert Sobhon
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  8. Pornanan Kueakhai
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Corresponding author

Correspondence to Pornanan Kueakhai.

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Changklungmoa, N., Chaithirayanon, K., Cheukamud, W. et al. Expression and characterization of glutathione peroxidase of the liver fluke, Fasciola gigantica. Parasitol Res 117, 3487–3495 (2018). https://doi.org/10.1007/s00436-018-6046-9

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  • DOI: https://doi.org/10.1007/s00436-018-6046-9

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