ArticleApplied and environmental microbiology2015
Scarless Genome Editing and Stable Inducible Expression Vectors for Geobacter sulfurreducens.
Article in Applied and environmental microbiology, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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30 citing papers in PubMed, 62 citations in OpenAlex.
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- Precise genome modification of agrobacterium based on one-step homologous recombination via endogenous transfer DNA.Acta biochimica Polonica · 2026Article
- Decoupled respiration in electro-active bacteria.Communications biology · 2025Article
- Evaluation of vectors for gene expression inApplied and environmental microbiology · 2025Article
- A widespread and ancient bacterial machinery assembles cytochrome OmcS nanowires essential for extracellular electron transfer.Cell chemical biology · 2025Article
- The role of CsrA in controls the extracellular electron transfer and biofilm production inFrontiers in microbiology · 2025Article
- A peptide-binding domain shared with an Antarctic bacterium facilitatesProceedings of the National Academy of Sciences of the United States of America · 2023Article
- Systematic Part Transfer by Extending a Modular Toolkit to Diverse Bacteria.ACS synthetic biology · 2023Article
- Conjugative plasmids inhibit extracellular electron transfer inFrontiers in microbiology · 2023Article
- Lack of Specificity inJournal of bacteriology · 2022Article
- Structure ofeLife · 2022Article
- Cryo-EM structure of an extracellular Geobacter OmcE cytochrome filament reveals tetrahaem packing.Nature microbiology · 2022Article
- The Signaling Pathway That cGAMP Riboswitches Found: Analysis and Application of Riboswitches to Study cGAMP Signaling inInternational journal of molecular sciences · 2022Article
- Article
- Evidence of a Streamlined Extracellular Electron Transfer Pathway from Biofilm Structure, Metabolic Stratification, and Long-Range Electron Transfer Parameters.Applied and environmental microbiology · 2021Article
- Engineering Biological Electron Transfer and Redox Pathways for Nanoparticle Synthesis.Bioelectricity · 2021Article
- Engineering Cooperation in an Anaerobic Coculture.Applied and environmental microbiology · 2021Article
- Engineering lithoheterotrophy in an obligate chemolithoautotrophic Fe(II) oxidizing bacterium.Scientific reports · 2021Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Metal reduction by members of the Geobacteraceae is encoded by multiple gene clusters, and the study of extracellular electron transfer often requires biofilm development on surfaces. Genetic tools that utilize polar antibiotic cassette insertions limit mutant construction and complementation. In addition, unstable plasmids create metabolic burdens that slow growth, and the presence of antibiotics such as kanamycin can interfere with the rate and extent of Geobacter biofilm growth. We report here genetic system improvements for the model anaerobic metal-reducing bacterium Geobacter sulfurreducens. A motile strain of G. sulfurreducens was constructed by precise removal of a transposon interrupting the fgrM flagellar regulator gene using SacB/sucrose counterselection, and Fe(III) citrate reduction was eliminated by deletion of the gene encoding the inner membrane cytochrome imcH. We also show that RK2-based plasmids were maintained in G. sulfurreducens for over 15 generations in the absence of antibiotic selection in contrast to unstable pBBR1 plasmids. Therefore, we engineered a series of new RK2 vectors containing native constitutive Geobacter promoters, and modified one of these promoters for VanR-dependent induction by the small aromatic carboxylic acid vanillate. Inducible plasmids fully complemented ΔimcH mutants for Fe(III) reduction, Mn(IV) oxide reduction, and growth on poised electrodes. A real-time, high-throughput Fe(III) citrate reduction assay is described that can screen numerous G. sulfurreducens strain constructs simultaneously and shows the sensitivity of imcH expression by the vanillate system. These tools will enable more sophisticated genetic studies in G. sulfurreducens without polar insertion effects or need for multiple antibiotics.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.