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The pGinger Family of Expression Plasmids
Allison N Pearson1,2,3, Mitchell G Thompson1,4, Liam D Kirkpatrick1,4
1Joint BioEnergy Institute, Emeryville, California, USA.
Microbiology Spectrum
|May 22, 2023
Summary
The pGinger plasmid suite offers 43 tools for precise gene expression in Gram-negative bacteria. These plasmids enable both constitutive and inducible control, expanding synthetic biology applications in diverse bacterial hosts.
Area of Science:
- Microbiology
- Synthetic Biology
- Molecular Biology
Background:
- Precise control of gene expression is fundamental for metabolic engineering and synthetic biology.
- Expanding synthetic biology beyond model organisms necessitates robust tools for diverse bacterial hosts.
Purpose of the Study:
- To develop and characterize a comprehensive suite of expression plasmids (pGinger) for precise constitutive and inducible gene expression.
- To provide a versatile set of tools for a wide range of Gram-negative bacteria, including nonmodel organisms.
Main Methods:
- Construction of 43 plasmids featuring synthetic constitutive promoters or inducible systems controlling red fluorescent protein (RFP) expression.
- Utilized broad-host-range origins (BBR1, RK2) and various antibiotic resistance markers (kanamycin, spectinomycin, gentamicin).
- Collected RFP expression and growth data in Escherichia coli and Pseudomonas putida.
Main Results:
- The pGinger suite includes 16 constitutive and 7 inducible expression vectors.
- Variants with different origins and selection markers were created for four inducible systems.
- Demonstrated functionality in model organisms like E. coli and P. putida.
Conclusions:
- The pGinger plasmid family provides a valuable resource for precise gene expression in a broad spectrum of Gram-negative bacteria.
- These tools will facilitate advancements in metabolic engineering and synthetic biology in nonmodel organisms.
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