With a daily production capacity of two grams of protein per egg, the chicken ovalbumin gene locus has the potential to synthesize large amounts of biopharmaceutical proteins into the eggs of transgenic hens. This phase I SBIR project will test the feasibility of using large expression constructs, based on the complete 200 kilobase chicken ovalbumin gene locus, to drive efficient expression of biopharmaceutical proteins including humanized monoclonal antibodies. The longer-term objectives of the program are to use these constructs in the creation of transgenic hens, in phase II that will produce biopharmaceutical proteins at mg/ml concentration in egg white. In this phase I proposal, we plan to create a complete 200 kilobase ovalbumin gene from two overlapping bacterial artificial chromosomes (BAC) by utilizing recA-assisted restriction endonuclease (RARE) cleavage techniques. Using similar techniques, we will then insert reporter and biopharmaceutical genes into various locations within the transcribed region of the ovalbumin gene. Finally, we will assay protein expression from these constructs using in vitro and in vivo assays. ? ?
