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>Effects of Plasmid Copy Number and Runaway Plasmid Replication on Overproduction and Excretion of β‐Lactamase fromEscherichiacoli
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Effects of Plasmid Copy Number and Runaway Plasmid Replication on Overproduction and Excretion of β‐Lactamase fromEscherichiacoli
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机译：Effects of Plasmid Copy Number and Runaway Plasmid Replication on Overproduction and Excretion of β‐Lactamase fromEscherichiacoli
AbstractRunaway plasmid replication can be used to increase target gene dosage and thereby overproduce proteins within the bacteriumEscherichia coli. However, the presence of excessive plasmid DNA often alters normal cell functions. High copy number plasmids with strong promoters place a severe metabolic burden on the cell, causing a decreased specific growth rate and changes in cell physiology. Induction of β‐lactamase synthesis from thetacpromoter on plasmid pKN causes runaway plasmid replication and excretion of β‐lactamase. Runaway plasmid relication results from readthrough oftacpromoter transcripts into the replication region of the plasmid. Both high plasmid copy numbers and a strong promoter (tac) are necessary to achieve the level of overproduction necessary for excretion of β‐lactamase, but high‐level target protein synthesis is detrimental to the cell. A derivative of pKN which is more easily regulated was constructed by adding the lacl gene to t
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