Ovalbumin locus BACs for expression of biotherapeutics

用于表达生物治疗药物的卵清蛋白位点 BAC

• 批准号: 6645148
• 负责人: MARK C. LEAVITT
• 金额: $ 8.15万
• 依托单位: SYNAGEVA BIOPHARMA CORPORATION
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2003-11-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6645148
• 关键词: artificial chromosomes; biotechnology; biotherapeutic agent; chickens; drug design /synthesis /production; egg /ovum; genetically modified animals; monoclonal antibody; ovalbumin; reporter genes; technology /technique development; transfection; transfection /expression vector

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：鸡卵蛋白基因基因座的每日生产能力为每卵两克蛋白质，有可能将大量的生物药物蛋白质合成到转基因母鸡的卵中。该阶段I SBIR项目将基于完整的200千生酶卵巢蛋白基因座基因座使用大型表达构建体的可行性，以驱动包括人源化单克隆抗体在内的生物药物蛋白的有效表达。该程序的长期目标是将这些构建体用于创建转基因母鸡，在II期中，将在蛋白中以mg/ml浓度产生生物药物蛋白质。在这一阶段的建议中，我们计划通过利用重新辅助的限制性核酸内切酶（稀有）裂解技术来从两个重叠的细菌人造染色体（BAC）中创建一个完整的200千生物卵巢基因。然后，我们将使用类似的技术将报告基因和生物药物基因插入卵巢蛋白基因转录区域内的各个位置。最后，我们将使用体外和体内测定法分析这些构建体的蛋白质表达。