Effects of Plasmid Copy Number and Runaway Plasmid Replication on Overproduction and Excretion of β‐Lactamase from Escherichia coli

A. Paul Togna; Michael L. Shuler; David B. Wilson

doi:10.1021/bp00019a005

journal article Jan 01, 1993

Effects of Plasmid Copy Number and Runaway Plasmid Replication on Overproduction and Excretion of β‐Lactamase from Escherichia coli

A. Paul Togna Michael L. Shuler

David B. Wilson

Biotechnology Progress Vol. 9 No. 1 pp. 31-39 · Wiley

View at Publisher Save 10.1021/bp00019a005

Abstract

AbstractRunaway plasmid replication can be used to increase target gene dosage and thereby overproduce proteins within the bacterium Escherichia 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 the tac promoter on plasmid pKN causes runaway plasmid replication and excretion of β‐lactamase. Runaway plasmid relication results from readthrough of tac promoter 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 the plasmid.

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References

Details

Published: Jan 01, 1993
Vol/Issue: 9(1)
Pages: 31-39
License: View

Authors

Cite This Article

A. Paul Togna, Michael L. Shuler, David B. Wilson (1993). Effects of Plasmid Copy Number and Runaway Plasmid Replication on Overproduction and Excretion of β‐Lactamase from Escherichia coli. Biotechnology Progress, 9(1), 31-39. https://doi.org/10.1021/bp00019a005

Effects of Plasmid Copy Number and Runaway Plasmid Replication on Overproduction and Excretion of β‐Lactamase from Escherichia coli

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