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

M. R. Mohamed, K. A. Shalaby, P. T. LoVerde, N. M. Abd Allah, A. M. Karim

Producción científica: Articlerevisión exhaustiva

5 Citas (Scopus)

Resumen

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.

Idioma originalEnglish (US)
Páginas (desde-hasta)1035-1042
Número de páginas8
PublicaciónParasitology Research
Volumen102
N.º5
DOI
EstadoPublished - abr 2008

ASJC Scopus subject areas

  • Insect Science
  • Infectious Diseases
  • General Veterinary
  • Parasitology

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