Abstract
| Original language | English |
|---|---|
| Article number | e19912 |
| Journal | P L o S One |
| Volume | 6 |
| Issue number | 5 |
| Number of pages | 6 |
| ISSN | 1932-6203 |
| DOIs | |
| Publication status | Published - 2011 |
| Externally published | Yes |
Bibliographical note
This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Keywords
- Agricultural and Biological Sciences (all)
- Biochemistry, Genetics and Molecular Biology (all)
- Medicine (all)
- Plasmid DNA
- Bacterial chromosome
- Bacterial genome
- Bacterial growth
- Bacterial reproduction
- Bacterium conjugation
- Controlled study
- DNA base composition
- DNA sequence
- Escherichia coli
- Gene dosage
- Gene frequency
- Gene synthesis
- Genetic load
- Horizontal gene transfer
- Nonhuman
- Nucleotide sequence
- PX1.0 plasmid
- Sequence analysis
- Bacteria (microorganisms)
- Enterobacteriaceae
- Conjugation, Genetic
- DNA, Bacterial
- Plasmids
Cite this
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Design and synthesis of a quintessential self-transmissible IncX1 plasmid, pX1.0. / Hansen, Lars Henrik; Bentzon-Tilia, Mikkel; Bentzon-Tilia, Sara; Norman, Anders; Rafty, Louise; Sørensen, Søren J.
In: P L o S One, Vol. 6, No. 5, e19912, 2011.Research output: Contribution to journal › Journal article › Research › peer-review
TY - JOUR
T1 - Design and synthesis of a quintessential self-transmissible IncX1 plasmid, pX1.0
AU - Hansen, Lars Henrik
AU - Bentzon-Tilia, Mikkel
AU - Bentzon-Tilia, Sara
AU - Norman, Anders
AU - Rafty, Louise
AU - Sørensen, Søren J.
N1 - This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2011
Y1 - 2011
N2 - DNA exchange in bacteria via conjugative plasmids is believed to be among the most important contributing factors to the rapid evolution- and diversification rates observed in bacterial species. The IncX1 plasmids are particularly interesting in relation to enteric bacteria, and typically carry genetic loads like antibiotic resistance genes and virulence factors. So far, however, a "pure" version of these molecular parasites, without genetic loads, has yet to be isolated from the environment. Here we report the construction of pX1.0, a fully synthesized IncX1 plasmid capable of horizontal transfer between different enteric bacteria. The designed pX1.0 sequence was derived from the consensus gene content of five IncX1 plasmids and three other, more divergent, members of the same phylogenetic group. The pX1.0 plasmid was shown to replicate stably in E. coli with a plasmid DNA per total DNA ratio corresponding to approximately 3-9 plasmids per chromosome depending on the growth phase of the host. Through conjugation, pX1.0 was able to self-transfer horizontally into an isogenic strain of E. coli as well as into two additional species belonging to the family Enterobacteriaceae. Our results demonstrate the immediate applicability of recent advances made within the field of synthetic biology for designing and constructing DNA systems, previously existing only in silica.
AB - DNA exchange in bacteria via conjugative plasmids is believed to be among the most important contributing factors to the rapid evolution- and diversification rates observed in bacterial species. The IncX1 plasmids are particularly interesting in relation to enteric bacteria, and typically carry genetic loads like antibiotic resistance genes and virulence factors. So far, however, a "pure" version of these molecular parasites, without genetic loads, has yet to be isolated from the environment. Here we report the construction of pX1.0, a fully synthesized IncX1 plasmid capable of horizontal transfer between different enteric bacteria. The designed pX1.0 sequence was derived from the consensus gene content of five IncX1 plasmids and three other, more divergent, members of the same phylogenetic group. The pX1.0 plasmid was shown to replicate stably in E. coli with a plasmid DNA per total DNA ratio corresponding to approximately 3-9 plasmids per chromosome depending on the growth phase of the host. Through conjugation, pX1.0 was able to self-transfer horizontally into an isogenic strain of E. coli as well as into two additional species belonging to the family Enterobacteriaceae. Our results demonstrate the immediate applicability of recent advances made within the field of synthetic biology for designing and constructing DNA systems, previously existing only in silica.
KW - Agricultural and Biological Sciences (all)
KW - Biochemistry, Genetics and Molecular Biology (all)
KW - Medicine (all)
KW - Plasmid DNA
KW - Bacterial chromosome
KW - Bacterial genome
KW - Bacterial growth
KW - Bacterial reproduction
KW - Bacterium conjugation
KW - Controlled study
KW - DNA base composition
KW - DNA sequence
KW - Escherichia coli
KW - Gene dosage
KW - Gene frequency
KW - Gene synthesis
KW - Genetic load
KW - Horizontal gene transfer
KW - Nonhuman
KW - Nucleotide sequence
KW - PX1.0 plasmid
KW - Sequence analysis
KW - Bacteria (microorganisms)
KW - Enterobacteriaceae
KW - Conjugation, Genetic
KW - DNA, Bacterial
KW - Plasmids
U2 - 10.1371/journal.pone.0019912
DO - 10.1371/journal.pone.0019912
M3 - Journal article
VL - 6
JO - P L o S One
JF - P L o S One
SN - 1932-6203
IS - 5
M1 - e19912
ER -