ArticleACS synthetic biology2017
A Family of Single Copy repABC-Type Shuttle Vectors Stably Maintained in the Alpha-Proteobacterium Sinorhizobium meliloti.
Article in ACS synthetic biology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 33 citations in OpenAlex.
- XanthoMoClo─A Robust Modular Cloning Genetic Toolkit for the GeneraACS synthetic biology · 2025Article
- Modular Low-Copy-Number Plasmid Vectors for Rhodobacterales with Extended Host Range in Alphaproteobacteria.ACS synthetic biology · 2024Article
- A MoClo-Compatible Toolbox of ECF Sigma Factor-Based Regulatory Switches for Proteobacterial Chassis.Biodesign research · 2024Article
- A Workflow for the Functional Characterization of Noncoding RNAs in Legume Symbiotic Bacteria.Methods in molecular biology (Clifton, N.J.) · 2024Article
- An Integrated Affinity Chromatography-Based Approach to Unravel the sRNA Interactome in Nitrogen-Fixing Rhizobia.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Genome-wide comparative analysis of clinical and environmental strains of the opportunistic pathogenFrontiers in microbiology · 2024Article
- Beyond the ABCs-Discovery of Three New Plasmid Types inMicroorganisms · 2022Article
- Genetic Engineering and Synthetic Genomics in Yeast to Understand Life and Boost Biotechnology.Bioengineering (Basel, Switzerland) · 2020Review
- Article
- SEVA 3.0: an update of the Standard European Vector Architecture for enabling portability of genetic constructs among diverse bacterial hosts.Nucleic acids research · 2020Article
- Design and Control of Extrachromosomal Elements inACS synthetic biology · 2019Article
- Reconsidering plasmid maintenance factors for computational plasmid design.Computational and structural biotechnology journal · 2019Review
- CRIMoClo plasmids for modular assembly and orthogonal chromosomal integration of synthetic circuits inJournal of biological engineering · 2019Article
- Designer Sinorhizobium meliloti strains and multi-functional vectors enable direct inter-kingdom DNA transfer.PloS one · 2019Article
- Horizontal operon transfer, plasmids, and the evolution of photosynthesis in Rhodobacteraceae.The ISME journal · 2018Article
- Seven-transmembrane receptor protein RgsP and cell wall-binding protein RgsM promote unipolar growth in Rhizobiales.PLoS genetics · 2018Article
- The Divided Bacterial Genome: Structure, Function, and Evolution.Microbiology and molecular biology reviews : MMBR · 2017Review
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
Abstract
A considerable share of bacterial species maintains segmented genomes. Plant symbiotic α-proteobacterial rhizobia contain up to six repABC-type replicons in addition to the primary chromosome. These low or unit-copy replicons, classified as secondary chromosomes, chromids, or megaplasmids, are exclusively found in α-proteobacteria. Replication and faithful partitioning of these replicons to the daughter cells is mediated by the repABC region. The importance of α-rhizobial symbiotic nitrogen fixation for sustainable agriculture and Agrobacterium-mediated plant transformation as a tool in plant sciences has increasingly moved biological engineering of these organisms into focus. Plasmids are ideal DNA-carrying vectors for these engineering efforts. On the basis of repABC regions collected from α-rhizobial secondary replicons, and origins of replication derived from traditional cloning vectors, we devised the versatile family of pABC shuttle vectors propagating in Sinorhizobium meliloti, related members of the Rhizobiales, and Escherichia coli. A modular plasmid library providing the elemental parts for pABC vector assembly was founded. The standardized design of these vectors involves five basic modules: (1) repABC cassette, (2) plasmid-derived origin of replication, (3) RK2/RP4 mobilization site (optional), (4) antibiotic resistance gene, and (5) multiple cloning site flanked by transcription terminators. In S. meliloti, pABC vectors showed high propagation stability and unit-copy number. We demonstrated stable coexistence of three pABC vectors in addition to the two indigenous megaplasmids in S. meliloti, suggesting combinability of multiple compatible pABC plasmids. We further devised an in vivo cloning strategy involving Cre/lox-mediated translocation of large DNA fragments to an autonomously replicating repABC-based vector, followed by conjugation-mediated transfer either to compatible rhizobia or E. coli.
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