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

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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.