<abstract xmlns="http://www.w3.org/1999/xhtml"><p>We demonstrate here for the first time the use of an IncP-1β plasmid, R751, as a gene capture vehicle for recombineering/conjugation strategies to clone large segments of bacterial genomes (20 – 100 + Kb). We designed R751 derivatives containing alternative markers for greater flexibility when using the R751 vehicle across different bacteria. These markers are removable if desired as part of the cloning procedure (with no extra steps needed). We demonstrated utility via cloning of 38 and 22 kb genomic segments from <italic>Salmonella enterica</italic> serovar Typhimurium and <italic>Escherichia coli</italic>, respectively. The plasmids expand the options available for use in recombineering/conjugation-based cloning applications.</p></abstract>

We demonstrate here for the first time the use of an IncP-1β plasmid, R751, as a gene capture vehicle for recombineering/conjugation strategies to clone large segments of bacterial genomes (20 – 100 + Kb). We designed R751 derivatives containing alternative markers for greater flexibility when using the R751 vehicle across different bacteria. These markers are removable if desired as part of the cloning procedure (with no extra steps needed). We demonstrated utility via cloning of 38 and 22 kb genomic segments from Salmonella enterica serovar Typhimurium and Escherichia coli, respectively. The plasmids expand the options available for use in recombineering/conjugation-based cloning applications.

Differentially Marked IncP-1β R751 Plasmids for Cloning via Recombineering and Conjugation

Polish Journal of Microbiology

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

We demonstrate here for the first time the use of an IncP-1β plasmid, R751, as a gene capture vehicle for recombineering/conjugation strategies to clone large...

Differentially Marked IncP-1β R751 Plasmids for Cloning via Recombineering and Conjugation

Article Category: short-communication

Published Online: Dec 05, 2019

Page range: 559 - 563

Received: Aug 20, 2019

Accepted: Oct 22, 2019

DOI: https://doi.org/10.33073/pjm-2019-052

Keywords
IncP-1, R751, FRT, FLP, Pdu, MCP

© 2019 Ashveen Bains and James W. Wilson et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Differentially Marked IncP-1β R751 Plasmids for Cloning via Recombineering and Conjugation

Article Category: short-communication

Published Online: Dec 05, 2019

Page range: 559 - 563

Received: Aug 20, 2019

Accepted: Oct 22, 2019

DOI: https://doi.org/10.33073/pjm-2019-052

KeywordsIncP-1, R751, FRT, FLP, Pdu, MCP

© 2019 Ashveen Bains and James W. Wilson et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Keywords
IncP-1, R751, FRT, FLP, Pdu, MCP