ArticleBMC microbiology2014
Construction of a mariner-based transposon vector for use in insertion sequence mutagenesis in selected members of the Rhizobiaceae.
Article in BMC microbiology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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26 citing papers in PubMed, 37 citations in OpenAlex.
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- Bacterial genome engineering using CRISPR-associated transposases.Nature protocols · 2024Review
- Rhizobium determinants of rhizosphere persistence and root colonization.The ISME journal · 2024Article
- CompoundingViruses · 2023Article
- A self-propagating, barcoded transposon system for the dynamic rewiring of genomic networks.Molecular systems biology · 2023Article
- Bacterial genome engineering using CRISPR RNA-guided transposases.bioRxiv : the preprint server for biology · 2023Article
- Elucidation of Essential Genes and Mutant Fitness during Adaptation toward Nitrogen Fixation Conditions in the Endophyte Azoarcus olearius BH72 Revealed by Tn-Seq.Microbiology spectrum · 2022Article
- PairedProceedings of the National Academy of Sciences of the United States of America · 2022Article
- TnMicrobiology spectrum · 2022Article
- Genome-Scale Mutational Analysis of Cathode-OxidizingFrontiers in microbiology · 2022Article
- Elucidating Essential Genes in Plant-Associated Pseudomonas protegens Pf-5 Using Transposon Insertion Sequencing.Journal of bacteriology · 2021Article
- Competition, Nodule Occupancy, and Persistence of Inoculant Strains: Key Factors in theFrontiers in plant science · 2021Review
- Lifestyle adaptations ofProceedings of the National Academy of Sciences of the United States of America · 2020Article
- Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing.Journal of visualized experiments : JoVE · 2020Article
- Application of Transposon Insertion Sequencing to Agricultural Science.Frontiers in plant science · 2020Review
- Rhizobia: from saprophytes to endosymbionts.Nature reviews. Microbiology · 2018Review
- Robustness encoded across essential and accessory replicons of the ecologically versatile bacterium Sinorhizobium meliloti.PLoS genetics · 2018Article
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2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe Rhizobiaceae family of Gram-negative bacteria often engage in symbiosis with plants of economic importance. Historically, genetic studies to identify the function of individual genes, and characterize the biology of these bacteria have relied on the use of classical transposon mutagenesis. To increase the rate of scientific discovery in the Rhizobiaceae there is a need to adapt high-throughput genetic screens like insertion sequencing for use in this family of bacteria. Here we describe a Rhizobiaceae compatible MmeI-adapted mariner transposon that can be used with insertion sequencing for high-throughput genetic screening.
resultsThe newly constructed mariner transposon pSAM_Rl mutagenized R. leguminosarum, S. meliloti, and A. tumefaciens at a high frequency. In R. leguminosarum, mutant pools were generated that saturated 88% of potential mariner insertions sites in the genome. Analysis of the R. leguminosarum transposon insertion sequencing data with a previously described hidden Markov model-based method resulted in assignment of the contribution of all annotated genes in the R. leguminosarum 3841 genome for growth on a complex medium. Good concordance was observed between genes observed to be required for growth on the complex medium, and previous studies.
conclusionsThe newly described Rhizobiaceaee compatible mariner transposon insertion sequencing vector pSAM_Rl has been shown to mutagenize at a high frequency and to be an effective tool for use in high-throughput genetic screening. The construction and validation of this transposon insertion sequencing tool for use in the Rhizobiziaceae will provide an opportunity for researchers in the Rhizobiaceae community to use high-throughput genetic screening, allowing for significant increase in the rate of genetic discovery, particularly given the recent release of genome sequences from many Rhizobiaceae strains.
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