ArticleBioscience of microbiota, food and health2013
The pyrE Gene as a Bidirectional Selection Marker in Bifidobacterium Longum 105-A.
Article in Bioscience of microbiota, food and health, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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15 citing papers in PubMed, 28 citations in OpenAlex.
- Machine Learning Reveals Quantitative Amino Acid Preferences in Bifidobacterium longum Growth.Microbial biotechnology · 2026Article
- Scarless gene disruption enabled by a dual-plasmid knockout platform in a clinical infant-derivedFrontiers in microbiology · 2025Article
- An improved temperature-sensitive shuttle vector system for scarless gene deletion in human-gut-associatediScience · 2024Article
- Genetic tools for the electrotrophApplied and environmental microbiology · 2024Article
- Recombinant Limosilactobacillus (Lactobacillus) delivering nanobodies against Clostridium perfringens NetB and alpha toxin confers potential protection from necrotic enteritis.MicrobiologyOpen · 2022Article
- Cell factory for γ-aminobutyric acid (GABA) production using Bifidobacterium adolescentis.Microbial cell factories · 2022Article
- A novel SfaNI-like restriction-modification system in Caldicellulosiruptor extents the genetic engineering toolbox for this genus.PloS one · 2022Article
- Identifying the essential nutritional requirements of the probiotic bacteria Bifidobacterium animalis and Bifidobacterium longum through genome-scale modeling.NPJ systems biology and applications · 2021Article
- A Resource for Cloning and Expression Vectors Designed for Bifidobacteria: Overview of Available Tools and Biotechnological Applications.Methods in molecular biology (Clifton, N.J.) · 2021Review
- Methionine utilization by bifidobacteria: possible existence of a reverse transsulfuration pathway.Bioscience of microbiota, food and health · 2021Article
- Methods for Isolation and Recovery of Bifidobacteria.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Bacterial Genetic Engineering by Means of Recombineering for Reverse Genetics.Frontiers in microbiology · 2020Review
- The essential genomic landscape of the commensal Bifidobacterium breve UCC2003.Scientific reports · 2017Article
- Capsular polysaccharide inhibits adhesion ofGut pathogens · 2017Article
- Complete Genome Sequence of Bifidobacterium longum 105-A, a Strain with High Transformation Efficiency.Genome announcements · 2014Article
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We constructed a deletion mutant of the pyrE gene in Bifidobacterium longum 105-A. A pyrE knockout cassette was cloned into pKKT427, a Bifidobacterium-Escherichia coli shuttle vector, and then introduced into B. longum 105-A by electroporation. The transformants were propagated and spread onto MRS plates containing 5-fluoroorotic acid (5-FOA) and uracil. 5-FOA-resistant mutants were obtained at a frequency of 4.7 × 10(-5) integrations per cell. To perform pyrE gene complementation, the pyrE gene was amplified by PCR and used to construct a complementation plasmid (pKKT427-pyrE (+)). B. longum 105-A ∆pyrE harboring this plasmid could not grow on MRS plates containing 5-FOA, uracil and spectinomycin. We also developed a chemically defined medium (bifidobacterial minimal medium; BMM) containing inorganic salts, glucose, vitamins, isoleucine and tyrosine for positive selection of pyrE transformants. B. longum 105-A ∆pyrE could not grow on BMM agar, but the same strain harboring pKKT427-pyrE (+) could. Thus, pyrE can be used as a counterselection marker in B. longum 105-A and potentially other Bifidobacterium species as well. We demonstrated the effectiveness of this system by constructing a knockout mutant of the xynF gene in B. longum 105-A by using the pyrE gene as a counterselection marker. This pyrE-based selection system will contribute to genetic studies of bifidobacteria.
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