ArticlePloS one2013
Overexpression of the tcp gene cluster using the T7 RNA polymerase/promoter system and natural transformation-mediated genetic engineering of Vibrio cholerae.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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12 citing papers in PubMed, 22 citations in OpenAlex.
- A Green Light-Regulated T7 RNA Polymerase Gene Expression System for Cyanobacteria.Marine biotechnology (New York, N.Y.) · 2021Article
- Interbacterial competition and anti-predatory behaviour of environmental Vibrio cholerae strains.Environmental microbiology · 2020Article
- Comparison of chitin-induced natural transformation in pandemic Vibrio cholerae O1 El Tor strains.Environmental microbiology · 2020Article
- A modular chromosomally integrated toolkit for ectopic gene expression in Vibrio cholerae.Scientific reports · 2020Article
- Neighbor predation linked to natural competence fosters the transfer of large genomic regions ineLife · 2019Article
- Ecological implications of gene regulation by TfoX and TfoY among diverse Vibrio species.Environmental microbiology · 2019Article
- Molecular insights into Vibrio cholerae's intra-amoebal host-pathogen interactions.Nature communications · 2018Article
- An intracellular replication niche for Vibrio cholerae in the amoeba Acanthamoeba castellanii.The ISME journal · 2016Article
- A Novel and Fast Purification Method for Nucleoside Transporters.Frontiers in molecular biosciences · 2016Article
- Regulatory elements involved in the expression of competence genes in naturally transformable Vibrio cholerae.BMC microbiology · 2014Article
- Glucose- but not rice-based oral rehydration therapy enhances the production of virulence determinants in the human pathogen Vibrio cholerae.PLoS neglected tropical diseases · 2014Article
- ComEA is essential for the transfer of external DNA into the periplasm in naturally transformable Vibrio cholerae cells.PLoS genetics · 2014Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
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Abstract
The human pathogen and aquatic bacterium Vibrio cholerae belongs to the group of naturally competent bacteria. This developmental program allows the bacterium to take up free DNA from its surrounding followed by a homologous recombination event, which allows integration of the transforming DNA into the chromosome. Taking advantage of this phenomenon we genetically engineered V. cholerae using natural transformation and FLP recombination. More precisely, we adapted the T7 RNA polymerase/promoter system in this organism allowing expression of genes in a T7 RNA polymerase-dependent manner. We naturally transformed V. cholerae by adding a T7-specific promoter sequence upstream the toxin-coregulated pilus (tcp) gene cluster. In a V. cholerae strain, which concomitantly produced the T7 RNA polymerase, this genetic manipulation resulted in the overexpression of downstream genes. The phenotypes of the strain were also in line with the successful production of TCP pili. This provides a proof-of-principle that the T7 RNA polymerase/promoter system is functional in V. cholerae and that genetic engineering of this organism by natural transformation is a straightforward and efficient approach.
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