ArticleNature protocols2011
Identifying microbial fitness determinants by insertion sequencing using genome-wide transposon mutant libraries.
Article in Nature protocols, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 123 papers.
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Who cites it
123 citing papers in PubMed.
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- Identification of immunogenic commensal antigens using phage display.Nature protocols · 2026Review
- Linking interpersonal differences in gut microbiota composition and drug biotransformation activity.bioRxiv : the preprint server for biology · 2026Article
- A commensal bacterium secretes effector proteins to establish population heterogeneity in the gut.Cell host & microbe · 2025Article
- Niche partitioning by resource size in the gut microbiome.bioRxiv : the preprint server for biology · 2025Article
- A human gut bacterium antagonizes neighboring bacteria by altering their protein-folding ability.Cell host & microbe · 2025Article
- Klebsiella pneumoniae employs a type VI secretion system to overcome microbiota-mediated colonization resistance.Nature communications · 2025Article
- Applying 3D cultures and high-throughput technologies to study host-pathogen interactions.Frontiers in immunology · 2025Review
- PIMMS-Dash: Accessible analysis, interrogation, and visualisation of high-throughput transposon insertion sequencing (TIS) data.Computational and structural biotechnology journal · 2024Article
- Genetic screen identifies cell wall enzyme is key for freshwater survival ofbioRxiv : the preprint server for biology · 2024Article
- InSeq analysis of definedmBio · 2024Article
- Review
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- Surface colonization bymBio · 2024Article
- A simple and cost-effective transformation system forFrontiers in microbiology · 2024Article
- Screening for Genes Involved in Outer Membrane Vesicle Biogenesis in Bacteroides thetaiotaomicron.Methods in molecular biology (Clifton, N.J.) · 2024Article
63 more citing papers are in PubMed but not listed here.
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Authors and funding
3 authors.
Funding
Abstract
Insertion sequencing (INSeq) is a method for determining the insertion site and relative abundance of large numbers of transposon mutants in a mixed population of isogenic mutants of a sequenced microbial species. INSeq is based on a modified mariner transposon containing MmeI sites at its ends, allowing cleavage at chromosomal sites 16-17 bp from the inserted transposon. Genomic regions adjacent to the transposons are amplified by linear PCR with a biotinylated primer. Products are bound to magnetic beads, digested with MmeI and barcoded with sample-specific linkers appended to each restriction fragment. After limited PCR amplification, fragments are sequenced using a high-throughput instrument. The sequence of each read can be used to map the location of a transposon in the genome. Read count measures the relative abundance of that mutant in the population. Solid-phase library preparation makes this protocol rapid (18 h), easy to scale up, amenable to automation and useful for a variety of samples. A protocol for characterizing libraries of transposon mutant strains clonally arrayed in a multiwell format is provided.
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