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Molecular cloning and phylogenetic analysis of Clonorchis sinensis elongation factor-1α

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Abstract

Elongation factor-1 (EF-1) plays a primary role in protein synthesis, e.g., in the regulation of cell growth, aging, motility, embryogenesis, and signal transduction. The authors identified a clone CsIH23 by immunoscreening a Clonorchis sinensis cDNA library. The cDNA of CsIH23 was found to have a putative open reading frame containing 461 amino acids with a predicted molecular mass of 50.5 kDa. Its polypeptide sequence was highly homologous with EF-1α of parasites and vertebrate animals. CsIH23 polypeptide contained three GTP/GDP-binding sites, one ribosome-binding domain, one actin-binding domain, one tRNA-binding domain, and two glyceryl-phosphoryl-ethanolamine attachment sites. Based on these primary and secondary structural similarities, it was concluded that CsIH23 cDNA encodes C. sinensis EF-1α (CsEF-1α). In a molecular phylogenic tree, CsEF-1α clustered with the EF-1α of helminthic parasites. Subsequently, CsEF-1α recombinant protein was bacterially overexpressed and purified by Ni-NTA affinity column chromatography. Immunoblotting using CsEF-1α recombinant protein produced positive signals for all serum samples tested from clonorchiasis, opisthorchiasis viverinii, and paragonimiasis westermani patients and normal healthy controls. These findings suggest that recombinant CsEF-1α is of limited usefulness as serodiagnostic antigen for clonorchiasis.

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Acknowledgment

This work was supported by the Korea Science and Engineering Foundation (KOSEF) grant funded by the Korean government (MOST; R01-2002-000-00261-0).

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Authors and Affiliations

  1. Department of Parasitology, Chung-Ang University College of Medicine, Seoul, 156-756, South Korea

    Jong Won Na & Sung-Jong Hong

  2. Department of Infection Biology, College of Medicine, Chungnam National University, Daejeon, 301-131, South Korea

    Tae Yun Kim

  3. Division of Malaria and Parasitic Diseases, National Institute of Health, Korea Centers for Disease Control and Prevention, Seoul, 122-701, South Korea

    Pyo Yun Cho

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  1. Tae Yun Kim
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  2. Pyo Yun Cho
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  3. Jong Won Na
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Correspondence to Sung-Jong Hong.

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Kim, T.Y., Cho, P.Y., Na, J.W. et al. Molecular cloning and phylogenetic analysis of Clonorchis sinensis elongation factor-1α. Parasitol Res 101, 1557–1562 (2007). https://doi.org/10.1007/s00436-007-0676-7

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  • DOI: https://doi.org/10.1007/s00436-007-0676-7

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