ArticleApplied and environmental microbiology1990
Thymidylate synthase gene from Lactococcus lactis as a genetic marker: an alternative to antibiotic resistance genes.
Article in Applied and environmental microbiology, 1990. 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, 46 citations in OpenAlex.
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- Article
- Insertional inactivation of determinants for Mg2+ and Co2+ transport as a tool for screening recombinant Lactococcus species clones.Applied and environmental microbiology · 2005Article
- Construction and characterization of a thyA mutant derived from cholera vaccine candidate IEM101.Molecular biotechnology · 2005Article
- ThyA as a selection marker in construction of food-grade host-vector and integration systems for Streptococcus thermophilus.Applied and environmental microbiology · 2004Article
- Gene cloning and expression and secretion of Listeria monocytogenes bacteriophage-lytic enzymes in Lactococcus lactis.Applied and environmental microbiology · 2000Article
- Genes expressed in Pseudomonas putida during colonization of a plant-pathogenic fungus.Applied and environmental microbiology · 2000Article
- Cloning, expression, and chromosomal stabilization of the Propionibacterium shermanii proline iminopeptidase gene (pip) for food-grade application in Lactococcus lactis.Applied and environmental microbiology · 1998Article
- Food-grade cloning and expression system for Lactococcus lactis.Applied and environmental microbiology · 1996Article
- Greenhouse and Field Evaluations of an Autoselective System Based on an Essential Thymidylate Synthase Gene for Improved Maintenance of Plasmid Vectors in Modified Rhizobium meliloti.Applied and environmental microbiology · 1995Article
- Physical and genetic map of the chromosome of Lactococcus lactis subsp. lactis IL1403.Journal of bacteriology · 1992Article
- Cloning and characterization of the thymidylate synthase gene from Lactococcus lactis subsp. lactis.Applied and environmental microbiology · 1990Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The potential of the thymidylate synthase thyA gene cloned from Lactococcus lactis subsp. lactis as a possible alternative selectable marker gene to antibiotic resistance markers has been examined. The thyA mutation is a recessive lethal one; thyA mutants cannot survive in environments containing low amounts of thymidine or thymine (such as Luria-Bertani medium) unless complemented by the thyA gene. The cloned thyA gene was strongly expressed in L. lactis subsp. lactis, Escherichia coli, Rhizobium meliloti, and a fluorescent Pseudomonas strain. In addition, when fused to a promoterless enteric lac operon, the thyA gene drove expression of the lac genes in a number of gram-negative bacteria. In transformation experiments with thyA mutants of E. coli and conjugation experiments with thyA mutants of R. meliloti, the lactococcal thyA gene permitted selection of transformants and transconjugants with the same efficiency as did genes for resistance to ampicillin, chloramphenicol, or tetracycline. Starting from the broad-host-range plasmid pGD500, a plasmid, designated pPR602, was constructed which is completely free of antibiotic resistance genes and has the lactococcal thyA gene fused to a promoterless lac operon. This plasmid will permit growth of thyA mutant strains in the absence of thymidine or thymine and has a number of unique restriction sites which can be used for cloning.
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