MOLECULAR-CLONING AND CHARACTERIZATION OF THE FILARIAL LIM DOMAIN PROTEINS AVL3-1 AND OVL3-1

A full-length cDNA of the filarial nematode Acanthocheilonema viteae w as isolated from a cDNA library of female worms, using a partial cDNA of the OvL3-1 gene of Onchocerca volvulus as a probe. The AvL3-1 cDNA contained an open reading frame which encoded for a protein with a the oretical molecular weight of 64 kDa. The deduced protein contained a p redicted signal sequence, a short repetitive motive of unknown functio n, and three LIM domains. The structure of the LIM domains was identic al to those of zyxin, a cptoskeleton-associated protein of chicken fib roblasts, suggesting that AvL3-1 has a similar role in filarial nemato des. The sequence information was used to isolate the homologous cDNA of O. volvulus by PCR from a cDNA library of female O. volvulus, which showed an overall identity of 76.9% to AvL3-1 on the protein level. A VL3-1 was expressed in Escherichia coli and the affinity-purified fusi on free protein was used to immunize jirds (Meriones unguiculatus). Im munization together with the adjuvant STP or with Freund's adjuvant in duced IgG and IgM antibody responses, but no significant protection ag ainst a challenge infection with L(3) of A. viteae, compared to approp riate control groups. (C) 1995 Academic Press, Inc.