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Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism

Wang, Zhijun and Xiang, Li and Shao, Junjie and Węgrzyn, Alicja and Węgrzyn, Grzegorz (2006) Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism. Microbial Cell Factories, 5 (1). 18 p.. ISSN 14752859


Official URL: http://dx.doi.org/10.1186/1475-2859-5-34


BACKGROUND: Although understanding of physiological interactions between plasmid DNA and its host is important for vector design and host optimization in many biotechnological applications, to our knowledge, global studies on plasmid-host interactions have not been performed to date even for well-characterized plasmids. RESULTS: Escherichia coli cells, either devoid of plasmid DNA or bearing plasmid pOri1 (with a single ColE1 replication origin) or plasmid pOri2 (with double ColE1 replication origins), were cultured in a chemostat. We used a combination of metabolic flux analysis, DNA microarray and enzyme activity analysis methods to explore differences in the metabolism between these strains. We found that the presence of plasmids significantly influenced various metabolic pathways in the host cells, e.g. glycolysis, the tricarboxylic acid (TCA) cycle and the pentose phosphate (PP) pathway. Expression of rpiA, a gene coding for ribose-5-phosphate isomerase A, was considerably decreased in E. coli carrying a high copy number plasmid relative to E. coli carrying a low copy number plasmid and plasmid-free E. coli. The rpiA gene was cloned into an expression vector to construct plasmid pETrpiA. Following induction of pETrpiA-bearing E. coli, which harbored either pOri1 or pOri2, with isopropyl-beta-D-thiogalactopyranoside (IPTG), the copy number of pOri1 and pOri2 was sigificantly higher than that measured in a host devoid of pETrpiA. CONCLUSION: The presence of plasmids can significantly influence some metabolic pathways in the host cell. We believe that the results of detailed metabolic analysis may be useful in optimizing host strains, vectors and cultivation conditions for various biotechnological purposes.

Item Type:Article
Subjects:Q Science > QH Natural history > QH301 Biology
Q Science > QH Natural history > QH426 Genetics
Q Science > QR Microbiology
Divisions:Laboratory of Molecular Biology (in Gdansk)
ID Code:931
Deposited By: PhD Piotr Golec
Deposited On:29 Sep 2015 13:07
Last Modified:29 Sep 2015 13:07

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