ArticleCurrent protocols in molecular biology2017
Mapping Transposon Insertions in Bacterial Genomes by Arbitrarily Primed PCR.
Article in Current protocols in molecular 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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17 citing papers in PubMed, 29 citations in OpenAlex.
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- An evolutionarily conserved metabolite inhibits biofilm formation in Escherichia coli K-12.Nature communications · 2024Article
- Genetic Underpinnings of Carotenogenesis and Light-Induced Transcriptome Remodeling in the Opportunistic PathogenPathogens (Basel, Switzerland) · 2023Article
- Human pathogenic bacteria on high-touch dry surfaces can be controlled by warming to human-skin temperature under moderate humidity.PloS one · 2023Article
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- InMut-finder: a software tool for insertion identification in mutagenesis using Nanopore long reads.BMC genomics · 2021Article
- Activation of the Type VI Secretion System in the Squid SymbiontJournal of bacteriology · 2021Article
- Identification of signaling pathways, matrix-digestion enzymes, and motility components controllingProceedings of the National Academy of Sciences of the United States of America · 2020Article
- The Polyphosphate Kinase of Escherichia coli Is Required for Full Production of the Genotoxin Colibactin.mSphere · 2020Article
- Programmable Gene Knockdown in Diverse Bacteria Using Mobile-CRISPRi.Current protocols in microbiology · 2020Article
- Genetic Manipulation of Vibrio fischeri.Current protocols in microbiology · 2020Article
- Quorum sensing controlseLife · 2018Article
- Efficient transposon mutagenesis mediated by an IPTG-controlled conditional suicide plasmid.BMC microbiology · 2018Article
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
Transposons can be used to easily generate and label the location of mutations throughout bacterial and other genomes. Transposon insertion mutants may be screened for a phenotype as individual isolates, or by selection applied to a pool of thousands of mutants. Identifying the location of a transposon insertion is critical for connecting phenotype to the genetic lesion. In this unit, we present an easy and detailed approach for mapping transposon insertion sites using arbitrarily-primed PCR (AP-PCR). Two rounds of PCR are used to (1) amplify DNA spanning the transposon insertion junction, and (2) increase the specific yield of transposon insertion junction fragments for sequence analysis. The resulting sequence is mapped to a bacterial genome to identify the site of transposon insertion. In this protocol, AP-PCR as it is routinely used to map sites of transposon insertion within Staphylococcus aureus, is used to illustrate the principle. Guidelines are provided for adapting this protocol for mapping insertions in other bacterial genomes. Mapping transposon insertions using this method is typically achieved in 2 to 3 days if starting from a culture of the transposon insertion mutant. © 2017 by John Wiley & Sons, Inc.
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