Design and synthesis of a quintessential self-transmissible IncX1 plasmid, pX1.0

Lars Henrik Hansen, Mikkel Bentzon-Tilia, Sara Bentzon-Tilia, Anders Norman, Louise Rafty, Søren J. Sørensen

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

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.
Original languageEnglish
Article numbere19912
JournalP L o S One
Volume6
Issue number5
Number of pages6
ISSN1932-6203
DOIs
Publication statusPublished - 2011
Externally publishedYes

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

Hansen, L. H., Bentzon-Tilia, M., Bentzon-Tilia, S., Norman, A., Rafty, L., & Sørensen, S. J. (2011). Design and synthesis of a quintessential self-transmissible IncX1 plasmid, pX1.0. P L o S One, 6(5), [e19912]. https://doi.org/10.1371/journal.pone.0019912
Hansen, Lars Henrik ; Bentzon-Tilia, Mikkel ; Bentzon-Tilia, Sara ; Norman, Anders ; Rafty, Louise ; Sørensen, Søren J. / Design and synthesis of a quintessential self-transmissible IncX1 plasmid, pX1.0. In: P L o S One. 2011 ; Vol. 6, No. 5.
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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",
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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 journalJournal articleResearchpeer-review

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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.

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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

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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

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M1 - e19912

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