We are interested in usage of C. elegans as system for expression of hookworm genes. Hookworms (Ancylostomatideae ) are economically and medically important blood feeding nematodes which cause intestinal infections in humans and domestic animals. It has been reported recently that above 1 billion of human beings are currently infected with these worms. The application of C. elegans as a heterologous host for hookworm genes is limited mainly by the lack of hookworm genomic data. For this purpose we have constructed cDNA and gDNA libraries from the Ancylostoma ceylanicum -important human/animal parasite related to C. elegans. The cDNA library is based on PCR mediated amplification of spliced leader of parasite cDNA reported previously ( Martin SA, Thompson FJ, Devaney E. Mol. Biochem. Parasitol. 1995 Mar;70(1-2):241-5.). PCR amplified cDNA has been gel fractionated and cloned. The most abundant library is available now from adult mixed sex A. ceylanicum (above 6000 clones), smaller library was constructed from infective L3 larvae. The large inserts gDNA libraries were constructed by ligation of PFGE fractionated Ancylostoma genomic DNA into the pBACe3.6 (provided by Pieter deJong), both digested with EcoRI. Ligation mixture was precipitated with ethanol in the presence of yeast tRNA prior electroporation into DH10B host. Both cDNA and gDNA libraries were arrayed in 96 well format, and stored at -70C. Randomly picked clones were sequenced from both ends, blastn and blastx searches were performed. For cDNA clones our sequencing results show clear similarities to C. elegans genes : gene F38E11.2
hsp-12.6; similar to Human Alpha crystallin B chain, gene C15C7.6; similar to syntaxin-6, gene T20F5.2; similar to the S25B family of peptidases. For BAC clones, end sequencing was also performed ( in collaboration with M. obocka, Institute of Biochemistry and Biophysics ) , however no significant similarities were found (yet!).