CLONING AND EXPRESSION OF THE HOOKWORM ANTICOAGULANT

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DESCRIPTION: Phase I of the planned program consists of taking a summer course in molecular biology and auditing five courses. Phases I and II will include different portions of the research program. Human hookworm infection is characterized by chronic GI bleeding and iron deficiency anemia. The adult worm facilitates blood feeding through secretion of a potent anticoagulant called the Hookworm Extrinsic Pathway Inhibitor (HEPI). The candidate has already isolated this novel 16 kDa protein from lysates of Ancylostoma duodenale. Synthetic oligonucleotides from amino acid sequences generated from tryptic digests of the protein have been used to generate and clone a PCR product containing a partial coding region sequence. The candidate plans to clone the full length gene from a cDNA library, to sequence this and to express recombinant HEPI in a bacterial or mammalian vector system. This expressed protein will be used to generate antibody for immunolocallization of the protein in the organism and for biochemical assays of protein activity.