An easily modifiable conjugative plasmid for studying horizontal gene transfer

Research output: Contribution to journalJournal articlepeer-review

Standard

An easily modifiable conjugative plasmid for studying horizontal gene transfer. / Wang, Qinqin; Olesen, Asmus Kalckar; Maccario, Lorrie; Madsen, Jonas Stenløkke.

In: Plasmid, Vol. 123-124, 102649, 2022.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Wang, Q, Olesen, AK, Maccario, L & Madsen, JS 2022, 'An easily modifiable conjugative plasmid for studying horizontal gene transfer', Plasmid, vol. 123-124, 102649. https://doi.org/10.1016/j.plasmid.2022.102649

APA

Wang, Q., Olesen, A. K., Maccario, L., & Madsen, J. S. (2022). An easily modifiable conjugative plasmid for studying horizontal gene transfer. Plasmid, 123-124, [102649]. https://doi.org/10.1016/j.plasmid.2022.102649

Vancouver

Wang Q, Olesen AK, Maccario L, Madsen JS. An easily modifiable conjugative plasmid for studying horizontal gene transfer. Plasmid. 2022;123-124. 102649. https://doi.org/10.1016/j.plasmid.2022.102649

Author

Wang, Qinqin ; Olesen, Asmus Kalckar ; Maccario, Lorrie ; Madsen, Jonas Stenløkke. / An easily modifiable conjugative plasmid for studying horizontal gene transfer. In: Plasmid. 2022 ; Vol. 123-124.

Bibtex

@article{800eef845f8843e4adf31179b4ed257b,
title = "An easily modifiable conjugative plasmid for studying horizontal gene transfer",
abstract = "Horizontal gene transfer is an important mechanism in bacterial evolution and can occur at striking frequencies when mediated by mobile genetic elements. Conjugative plasmids are mobile genetic elements that are main drivers of horizontal transfer and a major facilitator in the spread of antibiotic resistance genes. However, conjugative plasmid models that readily can be genetically modified with the aim to study horizontal transfer are not currently available. The aim of this study was to develop a conjugative plasmid model where the insertion of gene cassettes such as reporter genes (e.g., fluorescent proteins) or antibiotic resistance genes would be efficient and convenient. Here, we introduced a single attTn7 site into the conjugative broad-host-range IncP-1 plasmid pKJK5 in a non-disruptive manner. Furthermore, a version with lower transfer rate and a non-conjugative version of pKJK5-attTn7 were also constructed. The advantage of having the attTn7 sites is that genes of interest can be introduced in a single step with very high success rate using the Tn7 transposition system. In addition, larger genetic fragments can be inserted. To illustrate the efficacy of the constructed pKJK5 plasmids, they were complemented with sfGFP (a gene encoding superfolder green fluorescent protein) in addition to seven different β-lactamase genes representing the four known classes of β-lactamases.",
keywords = "Conjugative plasmid, Genetic engineering, Tn7 transposons, λ red recombination system",
author = "Qinqin Wang and Olesen, {Asmus Kalckar} and Lorrie Maccario and Madsen, {Jonas Stenl{\o}kke}",
note = "Publisher Copyright: {\textcopyright} 2022 The Authors",
year = "2022",
doi = "10.1016/j.plasmid.2022.102649",
language = "English",
volume = "123-124",
journal = "Plasmid",
issn = "0147-619X",
publisher = "Academic Press",

}

RIS

TY - JOUR

T1 - An easily modifiable conjugative plasmid for studying horizontal gene transfer

AU - Wang, Qinqin

AU - Olesen, Asmus Kalckar

AU - Maccario, Lorrie

AU - Madsen, Jonas Stenløkke

N1 - Publisher Copyright: © 2022 The Authors

PY - 2022

Y1 - 2022

N2 - Horizontal gene transfer is an important mechanism in bacterial evolution and can occur at striking frequencies when mediated by mobile genetic elements. Conjugative plasmids are mobile genetic elements that are main drivers of horizontal transfer and a major facilitator in the spread of antibiotic resistance genes. However, conjugative plasmid models that readily can be genetically modified with the aim to study horizontal transfer are not currently available. The aim of this study was to develop a conjugative plasmid model where the insertion of gene cassettes such as reporter genes (e.g., fluorescent proteins) or antibiotic resistance genes would be efficient and convenient. Here, we introduced a single attTn7 site into the conjugative broad-host-range IncP-1 plasmid pKJK5 in a non-disruptive manner. Furthermore, a version with lower transfer rate and a non-conjugative version of pKJK5-attTn7 were also constructed. The advantage of having the attTn7 sites is that genes of interest can be introduced in a single step with very high success rate using the Tn7 transposition system. In addition, larger genetic fragments can be inserted. To illustrate the efficacy of the constructed pKJK5 plasmids, they were complemented with sfGFP (a gene encoding superfolder green fluorescent protein) in addition to seven different β-lactamase genes representing the four known classes of β-lactamases.

AB - Horizontal gene transfer is an important mechanism in bacterial evolution and can occur at striking frequencies when mediated by mobile genetic elements. Conjugative plasmids are mobile genetic elements that are main drivers of horizontal transfer and a major facilitator in the spread of antibiotic resistance genes. However, conjugative plasmid models that readily can be genetically modified with the aim to study horizontal transfer are not currently available. The aim of this study was to develop a conjugative plasmid model where the insertion of gene cassettes such as reporter genes (e.g., fluorescent proteins) or antibiotic resistance genes would be efficient and convenient. Here, we introduced a single attTn7 site into the conjugative broad-host-range IncP-1 plasmid pKJK5 in a non-disruptive manner. Furthermore, a version with lower transfer rate and a non-conjugative version of pKJK5-attTn7 were also constructed. The advantage of having the attTn7 sites is that genes of interest can be introduced in a single step with very high success rate using the Tn7 transposition system. In addition, larger genetic fragments can be inserted. To illustrate the efficacy of the constructed pKJK5 plasmids, they were complemented with sfGFP (a gene encoding superfolder green fluorescent protein) in addition to seven different β-lactamase genes representing the four known classes of β-lactamases.

KW - Conjugative plasmid

KW - Genetic engineering

KW - Tn7 transposons

KW - λ red recombination system

U2 - 10.1016/j.plasmid.2022.102649

DO - 10.1016/j.plasmid.2022.102649

M3 - Journal article

C2 - 36100085

AN - SCOPUS:85138584451

VL - 123-124

JO - Plasmid

JF - Plasmid

SN - 0147-619X

M1 - 102649

ER -

ID: 322568662