![]() ![]() Among the various expression systems used, the temperature-inducible expression system has been beneficial due to the ease of induction and higher induction strength (Seeger et al. Several expression systems have been developed for the high-level expression of recombinant proteins in Escherichia coli (Makrides 1996). coli), we believe that such a systems-biological approach has several advantages over conventional experimental approaches and could be in principle extended to bigger genetic networks as well as other recombinant proteins and expression systems.Įscherichia coli has been used as an expression host for the production of a wide range of recombinant proteins (Baneyx 1999 Baneyx and Mujacic 2004). Although our results are specific to a recombinant protein (streptokinase) and the expression system ( E. In other words, manipulating the system properties resulting from the interaction of the components is better than manipulating the individual components. Analysis of the model simulations also shows that down regulation or selective redirection of the heat shock proteins could be a better way of manipulating the cellular stress response than overexpression or deletion. Detailed analysis of the parameter sensitivities of the model indicated that the level of free DnaK chaperone in the cell has the major effect on the productivity of recombinant streptokinase during temperature induction. ![]() The mechanistic mathematical model was validated with the experiments conducted on a σ 32 mutant strain. The model simulations were found to have a good qualitative agreement with the experimental results. Based on these investigations, a detailed mechanistic mathematical model was developed for the cellular response during the temperature-induced recombinant streptokinase production. The role played by the major molecular chaperone teams DnaK–DnaJ–GrpE and GroEL–GroES on the productivity of recombinant streptokinase was experimentally determined. To investigate the effect of the induced cellular response due to temperature up-shift a model-based approach is adopted. In this work, it was found that the temperature-inducible expression system for the production of soluble recombinant streptokinase in Escherichia coli resulted in a lower productivity compared to the chemically-induced system. The genetic networks involved in cellular stress response can have a critical effect on the productivity of recombinant proteins. Modeling and analysis of genetic networks have become increasingly important in the investigation of cellular processes. ![]()
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