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Principal Investigator: Dr. Theodore Clark
Contact Information: E-mail: tgc3@cornell.edu - Phone: 607-253-4042
Sponsor: Cornell Center for Advanced Technology
Title: A High-Copy Number Vector for Overexpression of Membrane Proteins
in Tetrahyemena
Annual Direct Cost: $50,000
Project Period: 07/01/05-06/30/06
Our laboratory has spent the past several years in efforts to develop Tetrahymena thermophila, a common pond-water ciliate, as a system for large-scale expression of recombinant proteins. Because of its unique biology, Tetrahymena is particularly well suited to the production of eucaryotic membrane and secretory proteins that are often difficult to express in more conventional systems such as E. coli and yeast. With our colleagues from the University of Georgia and the University of Rochester, methods for the stable transformation and inducible expression of foreign genes in this system have been developed resulting in one issued and two pending patents. Recently, this technology spawned a startup biotechnology company, Tetragenetics Inc. in Ithaca, New York. Tetragenetics intends to validate the commercial potential of this system by using it to express a variety of proteins of importance to human and animal health. These include potential vaccine candidates for the prevention of aquatic animal diseases. Of particular interest in this regard are two proteins, one a GPI-anchored surface antigen from Ichthyophthirius multifiliis, a protozoan pathogen that has major impact on commercial aquaculture worldwide, and the other a hemagglutinin from a cetacean morbillivirus of interest to the Navy. In this proposal, we will: 1) develop a high-copy number cloning vehicle (equivalent to an artificial chromosome that replicates to 18,000n) that should significantly enhance the expression of foreign genes in the Tetrahymena system, and 2) test this vector for its utility in overexpressing the viral and parasite proteins described above. Tetragenetics will commit matching funds for the proposed studies through in-kind support on a project to develop a non-marine mammalian model of the dolphin morbillivirus in which to test recombinant subunit vaccines against DMV.
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