Generation of a plasmid series for rapid sub-cloning and use in various Enterobacteriaceae
Date
2024-09-06
Authors
Braun, Hannah G.
Kanwal, Nabeela
Rivera Lopez, Luisa F.
Thomassin, Jenny-Lee
Journal Title
Journal ISSN
Volume Title
Publisher
Journal of Bioscience and Bioengineering
ORCID
Type
Article
Degree Level
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.
Description
Keywords
Plasmid construction, new tools, molecular biology
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
Degree
Department
Program
Advisor
Committee
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
Part Of
item.page.relation.ispartofseries
DOI
https://doi.org/10.1016/j.jbiosc.2024.08.006