ArticleApplied and environmental microbiology2001
Molecular characterization of a theta replication plasmid and its use for development of a two-component food-grade cloning system for Lactococcus lactis.
Article in Applied and environmental microbiology, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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19 citing papers in PubMed, 69 citations in OpenAlex.
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- Advances in Genetic Transformation of Lactic Acid Bacteria: Overcoming Barriers and Enhancing Plasmid Tools.International journal of molecular sciences · 2025Review
- Characterization of a novel theta-type replicon of indigenous plasmid pTE15 from Lactobacillus reuteri N16.BMC microbiology · 2022Article
- A host-vector toolbox for improved secretory protein overproduction in Bacillus subtilis.Applied microbiology and biotechnology · 2022Article
- Plasmid-Based Gene Expression Systems for Lactic Acid Bacteria: A Review.Microorganisms · 2022Review
- Plasmid Replicons for the Production of Pharmaceutical-Grade pDNA, Proteins and Antigens byInternational journal of molecular sciences · 2021Review
- Vector Development Timeline for Mucosal Vaccination and Treatment of Disease UsingFrontiers in microbiology · 2018Review
- Plasmids from Food Lactic Acid Bacteria: Diversity, Similarity, and New Developments.International journal of molecular sciences · 2015Review
- Characterization of plasmid pML21 of Enterococcus faecalis ML21 from koumiss.Current microbiology · 2013Article
- Article
- Adaptative potential of the Lactococcus lactis IL594 strain encoded in its 7 plasmids.PloS one · 2011Article
- Molecular characterization of the minimal replicon and the unidirectional theta replication of pSCM201 in extremely halophilic archaea.Journal of bacteriology · 2006Article
- Characterization of a galactokinase-positive recombinant strain of Streptococcus thermophilus.Applied and environmental microbiology · 2004Article
- Characterization of a theta-type plasmid from Lactobacillus sakei: a potential basis for low-copy-number vectors in lactobacilli.Applied and environmental microbiology · 2003Article
- Characterization of genes involved in the metabolism of alpha-galactosides by Lactococcus raffinolactis.Applied and environmental microbiology · 2003Article
- A food-grade approach for functional analysis and modification of native plasmids in Lactococcus lactis.Applied and environmental microbiology · 2003Article
- Novel food-grade plasmid vector based on melibiose fermentation for the genetic engineering of Lactococcus lactis.Applied and environmental microbiology · 2002Article
- Use of the alr gene as a food-grade selection marker in lactic acid bacteria.Applied and environmental microbiology · 2002Article
- Galactose and lactose genes from the galactose-positive bacterium Streptococcus salivarius and the phylogenetically related galactose-negative bacterium Streptococcus thermophilus: organization, sequence, transcription, and activity of the gal gene products.Journal of bacteriology · 2002Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
pCD4, a small, highly stable theta-replicating lactococcal plasmid, was used to develop a food-grade cloning system. Sequence analysis revealed five open reading frames and two putative cis-acting regions. None appears to code for undesirable phenotypes with regard to food applications. Functional analysis of the replication module showed that only the cis-acting ori region and the repB gene coding for the replication initiator protein were needed for the stable replication and maintenance of pCD4 derivatives in Lactococcus lactis. A two-component food-grade cloning system was derived from the pCD4 replicon. The vector pVEC1, which carries the functional pCD4 replicon, is entirely made up of L. lactis DNA and has no selection marker. The companion pCOM1 is a repB-deficient pCD4 derivative that carries an erythromycin resistance gene as a dominant selection marker. The pCOM1 construct can only replicate in L. lactis if trans complemented by the RepB initiator provided by pVEC1. Since only the cotransformants that carry both pVEC1 and pCOM1 can survive on plates containing erythromycin, pCOM1 can be used transiently to select cells that have acquired pVEC1. Due to the intrinsic incompatibility between these plasmids, pCOM1 can be readily cured from the cells grown on an antibiotic-free medium after the selection step. The system was used to introduce a phage resistance mechanism into the laboratory strain MG1363 of L. lactis and two industrial strains. The introduction of the antiphage barrier did not alter the wild-type plasmid profile of the industrial strains. The phenotype was stable after 100 generations and conferred an effective resistance phenotype against phages of the 936 and c2 species.
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