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. 1999 Jun 25;101(1-2):13-21.
doi: 10.1016/s0166-6851(99)00012-2.

Cloning and expression in Escherichia coli of a Fasciola hepatica gene encoding a calcium-binding protein

A D Ruiz de Eguino  1 , A Machín, R Casais, A M Castro, J A Boga, J M Martín-Alonso, F Parra
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Cloning and expression in Escherichia coli of a Fasciola hepatica gene encoding a calcium-binding protein

A D Ruiz de Eguino et al. Mol Biochem Parasitol. .

Abstract

A Fasciola hepatica cDNA clone of 994 bp was isolated from an adult worm cDNA expression library using a rabbit serum against the excretory-secretory antigens. The nucleotide sequence of the cDNA clone revealed the presence of an open reading frame of 572 bp which encoded a 22 kDa polypeptide (Fh22) showing putative EF-hand domains. This gene was expressed in Escherichia coli and the recombinant protein used for the production of specific antibodies. Immunoblotting studies using the anti-Fh22 serum showed the presence of a polypeptide of similar molecular mass in the excretory-secretory extract of the adult parasite. The recombinant Fh22 polypeptide showed calcium-dependent electrophoretic mobility (decreased with Ca2(+)-ions and increased with EGTA). The observed behaviour of recombinant Fh22 in gel filtration experiments also suggested calcium-induced conformational changes.

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