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Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism
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-ch...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2006
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1664580/ https://www.ncbi.nlm.nih.gov/pubmed/17112383 http://dx.doi.org/10.1186/1475-2859-5-34 |
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author | Wang, Zhijun Xiang, Li Shao, Junjie Węgrzyn, Alicja Węgrzyn, Grzegorz |
author_facet | Wang, Zhijun Xiang, Li Shao, Junjie Węgrzyn, Alicja Węgrzyn, Grzegorz |
author_sort | Wang, Zhijun |
collection | PubMed |
description | 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-β-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. |
format | Text |
id | pubmed-1664580 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2006 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-16645802006-11-30 Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism Wang, Zhijun Xiang, Li Shao, Junjie Węgrzyn, Alicja Węgrzyn, Grzegorz Microb Cell Fact Research 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-β-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. BioMed Central 2006-11-17 /pmc/articles/PMC1664580/ /pubmed/17112383 http://dx.doi.org/10.1186/1475-2859-5-34 Text en Copyright © 2006 Wang et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Wang, Zhijun Xiang, Li Shao, Junjie Węgrzyn, Alicja Węgrzyn, Grzegorz Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism |
title | Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism |
title_full | Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism |
title_fullStr | Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism |
title_full_unstemmed | Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism |
title_short | Effects of the presence of ColE1 plasmid DNA in Escherichia coli on the host cell metabolism |
title_sort | effects of the presence of cole1 plasmid dna in escherichia coli on the host cell metabolism |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1664580/ https://www.ncbi.nlm.nih.gov/pubmed/17112383 http://dx.doi.org/10.1186/1475-2859-5-34 |
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