Transcription from plasmid genes, macromolecular stability, and cell-specific productivity in Escherichia coli carrying copy number mutant plasmids.
Experiments were performed to evaluate, qualitatively and quantitatively, the adaptation of Escherichia coli to plasmid maintenance and cloned gene expression. Experimental findings indicate that the metabolic response to low plasmid levels is an increase of the biosynthetic capacity of both transcription and translation. At high copy number levels the gene-specific transcription rate continues to increase but the stability of plasmid-derived mRNA drops sharply. Protein levels are maintained, but translation efficiency decreases. These results indicate that cellular biosynthetic capacity may not be limiting productivity in recombinant systems. If macromolecular stability is the bottleneck, then current efforts to increase gene expression that focus on enhancing synthesis rates will be ineffective.
Peretti
S W
SW
Department of Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
Bailey
J E
JE
Lee
J J
JJ