Generation of a plasmid series for rapid sub-cloning and use in various Enterobacteriaceae
| dc.contributor.author | Braun, Hannah G. | |
| dc.contributor.author | Kanwal, Nabeela | |
| dc.contributor.author | Rivera Lopez, Luisa F. | |
| dc.contributor.author | Thomassin, Jenny-Lee | |
| dc.date.accessioned | 2024-09-28T19:03:19Z | |
| dc.date.available | 2024-09-28T19:03:19Z | |
| dc.date.issued | 2024-09-06 | |
| dc.description.abstract | Plasmids are molecular genetic tools used for trans-complementation and gene expression in bacteria. Challenges faced by researchers include limited repertoire of antibiotic resistance of plasmids, issues related to plasmid compatibility and restricted or incompatible multiple cloning sites when needing to change plasmid copy number to tune production of their protein of interest. In this study, a series of plasmids were generated with compatible multiple cloning sites and homologous DNA regions to allow for modular cloning for rapid exchange of antibiotic resistance and plasmid origin. Plasmids generated in this series have options for high, mid, and low plasmid copy number, and have either an integrated FLAG epitope in the multiple cloning site or possess an uninterrupted multiple cloning site with the option of using the common LacZ-based blue/white screening method. Low copy plasmids also have one of five antibiotic selection markers. To demonstrate functionality of these plasmids, a representative FLAG tagged protein and mCherry were cloned into the low copy plasmids and expressed in various bacteria belonging to the Enterobacteriaceae family. In conclusion, by creating a new plasmid series, we have expanded the toolkit of available molecular biology tools for bacterial work. | |
| dc.description.sponsorship | National Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant Saskatchewan Health Research Foundation establishment grant Canada Foundation for Innovation John R. Evans Leaders Fund Start-up funds from the College of Medicine at the University of Saskatchewan | |
| dc.description.version | Peer Reviewed | |
| dc.identifier.citation | Braun, H. G et al. Generation of a plasmid series for rapid sub-cloning and use in various Enterobacteriaceae. Journal of Bioscience and Bioengineering. https://doi.org/10.1016/j.jbiosc.2024.08.006 | |
| dc.identifier.doi | https://doi.org/10.1016/j.jbiosc.2024.08.006 | |
| dc.identifier.uri | https://hdl.handle.net/10388/16121 | |
| dc.language.iso | en | |
| dc.publisher | Journal of Bioscience and Bioengineering | |
| dc.subject | Plasmid construction | |
| dc.subject | new tools | |
| dc.subject | molecular biology | |
| dc.title | Generation of a plasmid series for rapid sub-cloning and use in various Enterobacteriaceae | |
| dc.type | Article |