Genetic Engineering of Proteins to Enhance Product Recovery

利用分子生物学赋予产品特性 其允许其从多组分细胞中选择性去除， 提取物在下游加工中具有巨大的益处， 生物技术产品，包括酶、激素和 抗体的 理论和过去的结果都表明， 向蛋白质中添加带电荷的尾部可以帮助选择性地 分离方法 该提议寻求将多肽尾融合到蛋白质中， 以提高蛋白质的回收率。 乳糖苷 已被选为模型蛋白质来实现这一建议。 工作计划包括以下内容： 1. 多肽尾部的设计。 2. 合成尾部的基因片段。 3. 将该片段克隆到β-半乳糖苷酶基因中。 4. 在E.杆菌 5. 评估结构和/或活动变化 目标蛋白质。 6. 通过沉淀回收融合蛋白， 两相水萃取 这份补助金申请是一个很好的例子， 我们希望在工程中看到的跨学科建议 董事会的生物技术计划。 主要研究者 被认为是有资格进行拟议的 research.

Using molecular biology to bestow a characteristic on a product that permits its selective removal from a multicomponent cellular extract can be of immense benefit in the downstream processing of the products of biotechnology, including enzymes, hormones and antibodies. Theory and past results both indicate that the addition of charged tails to proteins can aid the selectivity of separation methods. This proposal seeks to fuse polypeptide tails to proteins in order to enhance the recovery of proteins. Beta-galactosidase has been chosen as a model protein to carry out this proposal. The work plan includes the following: 1. Design of the polypeptide tail. 2. Synthesis of the gene segment for the tail. 3. Cloning this segment into the beta-galactosidase gene. 4. Expression of the gene in E. coli. 5. Assessment of structural and/or activity changes in the target protein. 6. Recovery of the fused protein by precipitation and two-phase aqueous extraction. This grant application is an excellent example of the type of cross-disciplinary proposal we wish to see in the Engineering Directorate's Biotechnology Program. The Principal Investigators are considered to be well qualified to carry out the proposed research.