ArticleApplied and environmental microbiology2013
A new large-DNA-fragment delivery system based on integrase activity from an integrative and conjugative element.
Article in Applied and environmental microbiology, 2013. 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, 31 citations in OpenAlex.
- Horizontal transfer of ICEJournal of bacteriology · 2026Article
- A comprehensive review of genomic-scale genetic engineering as a strategy to improve bacterial productivity.Microbiology (Reading, England) · 2025Review
- Advances in large-scale DNA engineering with the CRISPR system.Experimental & molecular medicine · 2025Review
- Validation of caprine H11 and the Rosa26 platform for transgene integration via CRISPR-based system: investigations on stable transgene expression and genetic biosafety.Functional & integrative genomics · 2025Article
- Isolation and genomic characterization of Psychrobacillus isolate L3 and bacteriophage Spoks: a new phage-host pair from Antarctic soil.BMC genomics · 2025Article
- It's complicated: relationships between integrative and conjugative elements and their bacterial hosts.Current opinion in microbiology · 2024Review
- TnSmu1 is a functional integrative and conjugative element in Streptococcus mutans that when expressed causes growth arrest of host bacteria.Molecular microbiology · 2022Article
- Biology and engineering of integrative and conjugative elements: Construction and analyses of hybrid ICEs reveal element functions that affect species-specific efficiencies.PLoS genetics · 2022Article
- Genetically engineered bacterium: Principles, practices, and prospects.Frontiers in microbiology · 2022Review
- Formation, Transmission, and Dynamic Evolution of a Multidrug-Resistant Chromosomally Integrated Plasmid inFrontiers in microbiology · 2022Article
- CRISPR RNA-guided integrases for high-efficiency, multiplexed bacterial genome engineering.Nature biotechnology · 2021Article
- Pass-back chain extension expands multimodular assembly line biosynthesis.Nature chemical biology · 2020Article
- The hidden life of integrative and conjugative elements.FEMS microbiology reviews · 2017Review
- Construction of an easy-to-use CRISPR-Cas9 system by patching a newly designed EXIT circuit.Journal of biological engineering · 2017Article
- New applications for phage integrases.Journal of molecular biology · 2014Review
- Biotechnological domestication of pseudomonads using synthetic biology.Nature reviews. Microbiology · 2014Review
- Engineering hydrogen gas production from formate in a hyperthermophile by heterologous production of an 18-subunit membrane-bound complex.The Journal of biological chemistry · 2014Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
During the past few decades, numerous plasmid vectors have been developed for cloning, gene expression analysis, and genetic engineering. Cloning procedures typically rely on PCR amplification, DNA fragment restriction digestion, recovery, and ligation, but increasingly, procedures are being developed to assemble large synthetic DNAs. In this study, we developed a new gene delivery system using the integrase activity of an integrative and conjugative element (ICE). The advantage of the integrase-based delivery is that it can stably introduce a large DNA fragment (at least 75 kb) into one or more specific sites (the gene for glycine-accepting tRNA) on a target chromosome. Integrase recombination activity in Escherichia coli is kept low by using a synthetic hybrid promoter, which, however, is unleashed in the final target host, forcing the integration of the construct. Upon integration, the system is again silenced. Two variants with different genetic features were produced, one in the form of a cloning vector in E. coli and the other as a mini-transposable element by which large DNA constructs assembled in E. coli can be tagged with the integrase gene. We confirmed that the system could successfully introduce cosmid and bacterial artificial chromosome (BAC) DNAs from E. coli into the chromosome of Pseudomonas putida in a site-specific manner. The integrase delivery system works in concert with existing vector systems and could thus be a powerful tool for synthetic constructions of new metabolic pathways in a variety of host bacteria.
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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.