Enhancement of Intracellular and Extracelluar Protein Production in Recombinant Yeast

维持高质粒稳定性和高效蛋白 分泌对于增强细胞内和 重组酵母胞外蛋白生产 啤酒。 本文概述的研究的一个主要目标是 建议研究质粒的稳定性和克隆的基因表达， 复杂介质 通过适当选择质粒， 无论使用何种营养培养基，都存在含有质粒的细胞。 详细的实验计划已被描述，重点是影响 培养基富集对克隆基因产物合成的影响。 的结果 这项研究将表明自动选择系统的潜力， 异源蛋白质的产生。 拟议工作的另一个主要重点是蛋白质分泌。 各种遗传和环境因素将被操纵， 确定它们对蛋白质分泌效率的影响。 在 特别是启动子强度和转录速率的影响， 蛋白质分泌的诱导和在复合培养基中的分泌将 追究 对于这两个研究领域，最初的工作将是实验性的。是 预计某些研究途径将服从动力学 建模

The maintenance of high plasmid stability and efficient protein secretion are important for the enhancement of intracellular and extracellular protein production in recombinant Saccharomyces cerevisiae. A major objective of the research outlined in this proposal is to study plasmid stability and cloned gene expression in complex media. With the proper choice of a plasmid, selection for plasmid-containing cells occurs regardless of the nutrient medium used. A detailed experimental plan has been described, focusing on the effects of medium enrichment on cloned gene product synthesis. The results of the research will indicate the potential of autoselection systems for the production of heterologous proteins. Another primary emphasis of the proposed work is protein secretion. Various genetic and environmental factors will be manipulated to determine their influence on the efficiency of protein secretion. In particular, the influence of promoter strength and transcription rate, the induction of protein secretion, and secretion in complex media will be investigated. For both areas of research, the initial work will be experimental. It is anticipated that certain avenues of research will be amenable to kinetic modeling.