Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Wang, Zhijun, Xiang, Li, Shao, Junjie, Węgrzyn, Alicja, Węgrzyn, Grzegorz
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2006
Materias:
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
_version_ 1782131070295080960
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
work_keys_str_mv AT wangzhijun effectsofthepresenceofcole1plasmiddnainescherichiacolionthehostcellmetabolism
AT xiangli effectsofthepresenceofcole1plasmiddnainescherichiacolionthehostcellmetabolism
AT shaojunjie effectsofthepresenceofcole1plasmiddnainescherichiacolionthehostcellmetabolism
AT wegrzynalicja effectsofthepresenceofcole1plasmiddnainescherichiacolionthehostcellmetabolism
AT wegrzyngrzegorz effectsofthepresenceofcole1plasmiddnainescherichiacolionthehostcellmetabolism