Determination of Genetic and Kinetic Parameters of Primary Importance to the Productivity of Recombinant Organisms

这项研究进行了一项基本的生化工程研究，以开发一种新的方法来确定对基因工程生物生产至关重要的动力学和遗传参数。这些重要参数包括：比产量、基因或质粒浓度、有质粒细胞的比例、有质粒细胞与无质粒细胞的生长比、特定的质粒损失率、相对质粒损失率、转录和翻译的集中速率以及基因的表达效率。目前，由于缺乏合理和系统的方法来准确确定决定和控制重组生物生产力的许多重要参数，这类数据库完全处于真空状态。鉴于这种情况，开发一种相对简单而又可靠的方法被认为是至关重要的，利用这种方法可以通过理论和实验研究来很好地表征和确定重组子的遗传和动力学参数。在前期研究的基础上，提出了一种能够准确确定这些参数的方法。根据这些参数，可以估计其他重要的参数，然后这些参数与环境和操作参数相结合，可以用于具有工业意义的重组发酵过程的设计、放大和优化。在这项研究中，所提出的理论方法将通过至少两个选定的“宿主细胞-载体”系统进行实验评估。根据实验结果，提出的确定重组生物遗传参数的方法将进一步完善或完善。

This investigation undertakes a fundamental biochemical engineering study to develop a novel methodology for determination of kinetic and genetic parameters of primary importance to the production of genetically engineered organisms. These important parameters include: specific production rate, gene or plasmid concentration, fraction of plasmid-harboring cells, growth ratio of plasmid- harboring cells to plasmid-free cells, specific plasmid loss rate, relative plasmid loss rate, lumped rate of transcription and translation, and gene expression efficiency. There is a total vacuum of the data base of this kind at the present time, because of total lack of rational and systematic methodology for accurate determination of many important parameters, which dictate and control the productivity of recombinant organisms. In view of this situation, it is considered critically important to develop a relatively simple and yet reliable method by which those genetic and kinetic parameters of recombinants can be well characterized and determined through both theoretical and experimental studies. Based on preliminary studies, one such method capable of determining accurately those parameters is proposed. From these parameters, other important parameters can be estimated and these parameters combined with the environmental and operational parameters can then be used for design, scale up, and optimization of recombinant fermentation processes of industrial importance. In this study, the proposed theoretical method will be evaluated experimentally by using at least two selected "host cell- vector" systems. Based on experimental results, the proposed method of determining the genetic parameters of recombinant organisms will be further improved or refined.