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Cloning and characterization of a cDNA fragment encoding a Schistosoma mansoni actin-binding protein (Smfilamin)

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Abstract

To identify vaccine candidates for Schistosoma mansoni, the IgG fraction of rabbit antiserum raised against immature female worms affinity purified over a NP-40 extract of 3-h schistosomula was used to immunoscreen a cercarial λgt11 cDNA library. One clone with a 1.5-kb cDNA insert revealed an encoded peptide of 479 amino acids, which bears homology to human actin-binding protein (ABP-280 = filamin). Northern blot analysis revealed a transcript of about 8.6 kb, indicating that the complete gene was not cloned. Overlapping clones, which encode a composite sequence of 983 amino acids (45% identity with filamin), were subsequently isolated from the cDNA library. The 1.5-kb insert was cloned into pGEX, overexpressed, and the 479 amino acid peptide purified. Western blot analysis using polyclonal antisera specific to the peptide identified a 280-kDa molecule in adult worm extracts. RT-PCR demonstrated that Smfilaimin is expressed in various stages. Immunofluorescence studies with specific antisera revealed a tegument-associated fluorescence in adult worms. IgG specific to the Smfilamin fragment showed 36.6% killing of schistosomules in an in vitro killing assay.

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Author notes

  1. P. T. LoVerde

    Present address: Departments of Biochemistry and Pathology, University of Texas Health Sciences, San Antonio, T, X, USA

Authors and Affiliations

  1. Department of Biochemistry, Faculty of Science, Ain Shams University, Cairo, Egypt

    M. R. Mohamed, K. A. Shalaby, N. M. Abd Allah & A. M. Karim

  2. Department of Microbiology, State University of New York, Buffalo, NY, USA

    P. T. LoVerde

Authors
  1. M. R. Mohamed
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  2. K. A. Shalaby
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  3. P. T. LoVerde
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  4. N. M. Abd Allah
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  5. A. M. Karim
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Corresponding author

Correspondence to A. M. Karim.

Additional information

This research was supported by grants from the NIH (AI18867) and the Schistosome Research Project (01-01-06). The nucleotide sequences reported in this paper were deposited in the GenBank (accession #AY463158). The experiments performed herein comply with the current laws of Egypt and the United States of America.

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Mohamed, M.R., Shalaby, K.A., LoVerde, P.T. et al. Cloning and characterization of a cDNA fragment encoding a Schistosoma mansoni actin-binding protein (Smfilamin). Parasitol Res 102, 1035–1042 (2008). https://doi.org/10.1007/s00436-007-0872-5

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  • DOI: https://doi.org/10.1007/s00436-007-0872-5

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