ArticleApplied and environmental microbiology2002
Efficient insertion mutagenesis strategy for bacterial genomes involving electroporation of in vitro-assembled DNA transposition complexes of bacteriophage mu.
Article in Applied and environmental microbiology, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 67 citations in OpenAlex.
- Tn5 DNA Transposase in Multi-Omics Research.Methods and protocols · 2023Review
- The DNA polymerase of bacteriophage YerA41 replicates its T-modified DNA in a primer-independent manner.Nucleic acids research · 2022Article
- Screening of Bacteriophage Encoded Toxic Proteins with a Next Generation Sequencing-Based Assay.Viruses · 2021Article
- Discovery of Three Toxic Proteins ofViruses · 2020Article
- A Toxicity Screening Approach to Identify Bacteriophage-Encoded Anti-Microbial Proteins.Viruses · 2019Article
- Targeted DNA transposition in vitro using a dCas9-transposase fusion protein.Nucleic acids research · 2019Article
- Transposase subunit architecture and its relationship to genome size and the rate of transposition in prokaryotes and eukaryotes.Nucleic acids research · 2018Article
- Mu transpososome activity-profiling yields hyperactive MuA variants for highly efficient genetic and genome engineering.Nucleic acids research · 2018Article
- Mu-driven transposition of recombinant mini-Mu unit DNA in the Corynebacterium glutamicum chromosome.Applied microbiology and biotechnology · 2018Article
- DNA Transposition at Work.Chemical reviews · 2016Review
- MuA-mediated in vitro cloning of circular DNA: transpositional autointegration and the effect of MuB.Molecular genetics and genomics : MGG · 2016Article
- Generation of comprehensive transposon insertion mutant library for the model archaeon, Haloferax volcanii, and its use for gene discovery.BMC biology · 2014Article
- An assay to monitor the activity of DNA transposition complexes yields a general quality control measure for transpositional recombination reactions.Mobile genetic elements · 2014Article
- Transposable Phage Mu.Microbiology spectrum · 2014Review
- Article
- Application of the bacteriophage Mu-driven system for the integration/amplification of target genes in the chromosomes of engineered Gram-negative bacteria--mini review.Applied microbiology and biotechnology · 2011Review
- Universal platform for quantitative analysis of DNA transposition.Mobile DNA · 2010Article
- Generation of single-copy transposon insertions in Clostridium perfringens by electroporation of phage mu DNA transposition complexes.Applied and environmental microbiology · 2009Article
- Bacteriophage Mu integration in yeast and mammalian genomes.Nucleic acids research · 2008Article
- Isolation and characterization of biofilm formation-defective mutants of Staphylococcus aureus.Infection and immunity · 2007Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
An efficient insertion mutagenesis strategy for bacterial genomes based on the phage Mu DNA transposition reaction was developed. Incubation of MuA transposase protein with artificial mini-Mu transposon DNA in the absence of divalent cations in vitro resulted in stable but inactive Mu DNA transposition complexes, or transpososomes. Following delivery into bacterial cells by electroporation, the complexes were activated for DNA transposition chemistry after encountering divalent metal ions within the cells. Mini-Mu transposons were integrated into bacterial chromosomes with efficiencies ranging from 10(4) to 10(6) CFU/microg of input transposon DNA in the four species tested, i.e., Escherichia coli, Salmonella enterica serovar Typhimurium, Erwinia carotovora, and Yersinia enterocolitica. Efficiency of integration was influenced mostly by the competence status of a given strain or batch of bacteria. An accurate 5-bp target site duplication flanking the transposon, a hallmark of Mu transposition, was generated upon mini-Mu integration into the genome, indicating that a genuine DNA transposition reaction was reproduced within the cells of the bacteria studied. This insertion mutagenesis strategy for microbial genomes may be applicable to a variety of organisms provided that a means to introduce DNA into their cells is available.
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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.