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Flow Cytometric Analysis of Metabolic Stress Effects Due to Recombinant Plasmids and Proteins in Escherichia coli Production Strains

✍ Scribed by Elena Soriano; Nicole Borth; Hermann Katinger; Diethard Mattanovich

Publisher: Elsevier Science
Year: 1999
Tongue: English
Weight: 427 KB
Volume: 1
Category: Article
ISSN: 1096-7176
DOI: 10.1006/mben.1999.0128

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✦ Synopsis

Overexpression of recombinant proteins in Escherichia coli often leads to a severe growth retardation of the host cells. Using flow cytometry, we analyzed the temporal development of the cellular content of DNA, total protein, and the recombinant product (human superoxide dismutase) in different strains. In cells carrying plasmids utilizing the phage T7 promoter ,10 (pET vectors), induction with IPTG leads to an increase in protein content and size, an increase and a wide spreading of DNA content distribution, and a termination of cell division. These effects occurred with pET plasmids with or without an insert, but not with another plasmid which utilizes the tac promoter.

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