ArticleApplied and environmental microbiology1990
Cloning and characterization of the thymidylate synthase gene from Lactococcus lactis subsp. lactis.
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 14 papers.
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Who cites it
14 citing papers in PubMed, 38 citations in OpenAlex.
- Metagenome Mining Reveals Hidden Genomic Diversity of Pelagimyophages in Aquatic Environments.mSystems · 2020Article
- Article
- 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
- Bacteriophage resistance of a deltathyA mutant of Lactococcus lactis blocked in DNA replication.Applied and environmental microbiology · 2002Article
- Identification of a thymidylate synthase gene within the genome of Chilo iridescent virus.Virus genes · 1998Article
- A general method for the consecutive integration of single copies of a heterologous gene at multiple locations in the Bacillus subtilis chromosome by replacement recombination.Applied and environmental microbiology · 1995Article
- 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 Lactococcus lactis subsp. cremoris MG1363 chromosome: comparison with that of Lactococcus lactis subsp. lactis IL 1403 reveals a large genome inversion.Journal of bacteriology · 1995Article
- Cloning and molecular analysis of the dihydrofolate reductase gene from Lactococcus lactis.Applied and environmental microbiology · 1995Article
- Expression, purification, and characterization of thymidylate synthase from Lactococcus lactis.Protein science : a publication of the Protein Society · 1994Article
- Physical and genetic map of the chromosome of Lactococcus lactis subsp. lactis IL1403.Journal of bacteriology · 1992Article
- New nucleotide sequence data on the EMBL File Server.Nucleic acids research · 1990Article
- Thymidylate synthase gene from Lactococcus lactis as a genetic marker: an alternative to antibiotic resistance genes.Applied and environmental microbiology · 1990Article
Corrections and comments
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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 thymidylate synthase (thyA) gene has been isolated from Lactococcus lactis subsp. lactis. The cloned gene was strongly expressed in Escherichia coli both in vivo and in vitro (maxicells and cell-free transcription and translation systems) and complemented E. coli thyA mutants. DNA-DNA hybridizations demonstrated that the thyA gene is encoded by the chromosome of L. lactis subsp. lactis. By sequential deletion of DNA outside the complementing region, the thyA gene was localized to a 1.1-kilobase DNA fragment. The nucleotide sequence of the lactococcal thyA gene was determined by the dideoxy-chain termination technique. The derived amino acid sequence indicated a protein size of 32,580 daltons, which is in good agreement with results obtained from maxicell and in vitro transcription and translation experiments. The primary sequence is homologous to 12 other thyA proteins from a variety of other organisms. Upstream from the structural gene, -10 and -35 promoter sequences which were almost canonical sigma-70 promoter sequences were identified, which may explain the strong expression of the thyA gene observed in E. coli. An A-T-rich sequence characteristic of gram-positive promoters was also noted adjacent to the -35 region. The thyA gene has potential as a marker for plasmid maintenance and selection in food systems.
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