Fine regulation of cI857-controlled gene expression in continuous culture of recombinant Escherichia coli by temperature

The expression at different temperatures of the lacZ gene, which is controlled by the lambda pL and pR tandem promoters and the cI857 temperature-sensitive repressor, was studied in Escherichia coli continuous cultures. At temperatures between 30 and 42 degrees C, beta-galactosidase activity behaved according to an exponential equation. By inducing a culture at a temperature within this range, predefined, nearly constant submaximal levels of gene expression and recombinant product yield can be obtained.

[1]  M. Lieb.  Studies of heat-inducible lambda bacteriophage. I. Order of genetic sites and properties of mutant prophages. , 1966, Journal of molecular biology.

[2]  Jeffrey H. Miller.  Experiments in molecular genetics , 1972 .

[3]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[4]  T. Platt,et al.  Maximizing gene expression from plasmid vectors containing the lambda PL promoter: strategies for overproducing transcription termination factor rho. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[5]  M. Rhodes,et al.  Temperature-induced synthesis of recombinant proteins , 1986 .

[6]  H. Blöcker,et al.  Inducible expression vectors incorporating the Escherichia coli atpE translational initiation region. , 1987, Gene.

[7]  K. Turner,et al.  Effect of induction temperature on the production of malaria antigens in recombinant E. coli , 1989, Biotechnology and bioengineering.

[8]  M. Bina,et al.  Temperature-mediated regulation and downstream inducible selection for controlling gene expression from the bacteriophage lambda pL promoter. , 1990, Gene.

[9]  A. Hortaçsu,et al.  Optimal Temperature Control Policy for a Two‐Stage Recombinant Fermentation Process , 1990, Biotechnology progress.

[10]  C. Schein,et al.  Optimizing protein folding to the native state in bacteria. , 1991, Current opinion in biotechnology.

[11]  Induction of recombinant gene expression in Escherichia coli using an alkaline pH shift. , 1991, Gene.

[12]  S. Enfors,et al.  Factors influencing inclusion body formation in the production of a fused protein in Escherichia coli , 1991, Applied and environmental microbiology.

[13]  A. Villaverde,et al.  Enhanced production of pL-controlled recombinant proteins and plasmid stability in Escherichia coli RecA+ strains. , 1993, Journal of biotechnology.

[14]  By-product formation , degradation and intracellular accumulation of recombinant protein , .