ArticleApplied and environmental microbiology2004
ThyA as a selection marker in construction of food-grade host-vector and integration systems for Streptococcus thermophilus.
Article in Applied and environmental microbiology, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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9 citing papers in PubMed.
- The Chromosomal Gene Manipulation Method for Lactobacillus delbrueckii subsp. bulgaricus Using a Conjugative Shuttle Vector pGMβ1.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Advances in Genetic Tools and Their Application inFoods (Basel, Switzerland) · 2023Review
- Effect of sugar transporter on galactose utilization inFrontiers in microbiology · 2023Article
- Novel shuttle vector pGMβ1 for conjugative chromosomal manipulation ofBioscience of microbiota, food and health · 2022Article
- Article
- Challenges and Advances for Genetic Engineering of Non-model Bacteria and Uses in Consolidated Bioprocessing.Frontiers in microbiology · 2017Review
- Genetic dissection of the polyoxin building block-carbamoylpolyoxamic acid biosynthesis revealing the "pathway redundancy" in metabolic networks.Microbial cell factories · 2013Article
- Construction and application of a food-grade expression system for Lactococcus lactis.Molecular biotechnology · 2013Article
- Development of a double-crossover markerless gene deletion system in Bifidobacterium longum: functional analysis of the α-galactosidase gene for raffinose assimilation.Applied and environmental microbiology · 2012Article
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3 authors.
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Abstract
We constructed food-grade host-vector and integration systems for Streptococcus thermophilus by using a thymidylate synthase gene (thyA) as the selection marker. Two thyA genes, thyA(St) and thyA(Lb), were cloned from S. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, respectively. Thymidine-requiring mutants of S. thermophilus were obtained after successive cultures in the presence of trimethoprim, and one of them, TM1-1, was used as the host. Food-grade vectors were constructed by using either thyA(St) or thyA(Lb) as the selection marker. Transformants of TM1-1 created by using these vectors were selected for thymidine autotrophy as efficiently as for erythromycin resistance. By using the host-vector system developed in this way, a foreign amylase gene (amyA) was expressed in TM1-1 and was also integrated into the chromosome by use of a temperature-sensitive integration vector constructed with thyA(Lb) as the selection marker via a double-crossover event. The results obtained show that thyA is an efficient and safe selection marker for S. thermophilus that is suitable for food applications.
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