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Updated: Apr 18, 2026
Testing the Role of Multicopy Plasmids in the Evolution of Antibiotic Resistance
Published on: May 2, 2018
Construction of the recombinant broad-host-range plasmids providing their bacterial hosts arsenic resistance and
Lukasz Drewniak1, Martyna Ciezkowska1, Monika Radlinska2
1Laboratory of Environmental Pollution Analysis, Faculty of Biology, University of Warsaw, Miecznikowa 1, 02-096 Warsaw, Poland.
Abstract:
The plasmid pSinA of Sinorhizobium sp. M14 was used as a source of functional phenotypic modules, encoding proteins involved in arsenite oxidation and arsenic resistance, to obtain recombinant broad-host-range plasmids providing their bacterial hosts arsenic resistance and arsenite oxidative ability. An arsenite oxidation module was cloned into pBBR1MCS-2 vector yielding plasmid vector pAIO1, while an arsenic resistance module was cloned into pCM62 vector yielding plasmid pARS1. Both plasmid constructs were introduced (separately and together) into the cells of phylogenetically distant (representing Alpha-, Beta-, and Gammaproteobacteria) and physiologically diversified (unable to oxidize arsenite and susceptible/resistant to arsenite and arsenate) bacteria. Functional analysis of the modified strains showed that: (i) the plasmid pARS1 can be used for the construction of strains with an increased resistance to arsenite [up to 20mM of As(III), (ii) the presence of the plasmid pAIO1 in bacteria previously unable to oxidize As(III) to As(V), contributes to the acquisition of arsenite oxidation abilities by these cells, (iii) the highest arsenite utilization rate are observed in the culture of strains harbouring both the plasmids pAIO1 and pARS1, (iv) the strains harbouring the plasmid pAIO1 were able to grow on arsenic-contaminated mine waters (∼ 3.0 mg As L(-1)) without any supplementation.
Insights
This study engineered bacteria with new plasmids for arsenic resistance and oxidation. These modified microbes can now thrive in arsenic-contaminated environments, offering potential bioremediation solutions.
Area of Science:
- Environmental Microbiology
- Biotechnology
- Genetic Engineering
Background:
- Arsenic contamination poses significant environmental and health risks.
- Microbial arsenic oxidation and resistance are crucial for bioremediation strategies.
- Sinorhizobium sp. M14 harbors genetic modules for arsenic detoxification.
Purpose of the Study:
- To develop recombinant broad-host-range plasmids for bacterial arsenic resistance and arsenite oxidation.
- To confer arsenic resistance and arsenite oxidative abilities to diverse bacterial hosts.
- To evaluate the efficacy of engineered plasmids in arsenic-contaminated environments.
Main Methods:
- Cloning of arsenite oxidation and arsenic resistance modules from Sinorhizobium sp. M14 into pBBR1MCS-2 (pAIO1) and pCM62 (pARS1) vectors, respectively.
- Introduction of engineered plasmids into phylogenetically diverse bacterial hosts (Alpha-, Beta-, and Gammaproteobacteria).
- Functional analysis of modified strains for arsenite oxidation, arsenic resistance, and growth in contaminated mine water.
Main Results:
- Plasmid pARS1 conferred increased arsenite resistance (up to 20mM As(III)).
- Plasmid pAIO1 enabled previously non-oxidizing bacteria to oxidize arsenite (As(III)) to arsenate (As(V)).
- Co-harboring both plasmids (pAIO1 and pARS1) resulted in the highest arsenite utilization rates.
- Strains with pAIO1 demonstrated growth in arsenic-contaminated mine water (∼3.0 mg As L⁻¹) without supplementation.
Conclusions:
- Recombinant plasmids pAIO1 and pARS1 effectively transfer arsenite oxidation and arsenic resistance traits to diverse bacterial hosts.
- Engineered bacteria exhibit enhanced capabilities for arsenic detoxification and utilization.
- These findings support the development of microbial-based solutions for arsenic bioremediation.
