ArticleApplied and environmental microbiology2007
Acetamide selection of Kluyveromyces lactis cells transformed with an integrative vector leads to high-frequency formation of multicopy strains.
Article in Applied and environmental microbiology, 2007. 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, 42 citations in OpenAlex.
- Creating a large designer cellulosome in yeast to boost ethanol production.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Proteolytic stabilization of a spider venom peptide results in an orally active bioinsecticide.Pest management science · 2025Article
- A convenient broad-host counterselectable system endowing rapid genetic manipulations of Kluyveromyces lactis and other yeast species.Microbial cell factories · 2024Article
- Identification of a Yarrowia lipolytica acetamidase and its use as a yeast genetic marker.Microbial cell factories · 2020Article
- Constructing a yeast to express the largest cellulosome complex on the cell surface.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- A Novel Regulated Hybrid Promoter That Permits Autoinduction of Heterologous Protein Expression in Kluyveromyces lactis.Applied and environmental microbiology · 2019Article
- Immunological Comparison of Native and Recombinant Hen's Egg Yolk Allergen, Chicken Serum Albumin (Gal d 5), Produced in Kluveromyces lactis.Nutrients · 2018Article
- Article
- Heterologous expression of an α-amylase inhibitor from common bean (Phaseolus vulgaris) in Kluyveromyces lactis and Saccharomyces cerevisiae.Microbial cell factories · 2017Article
- Development of a Candida glabrata dominant nutritional transformation marker utilizing the Aspergillus nidulans acetamidase gene (amdS).FEMS yeast research · 2016Article
- Metabolic engineering of Kluyveromyces lactis for L-ascorbic acid (vitamin C) biosynthesis.Microbial cell factories · 2013Article
- amdSYM, a new dominant recyclable marker cassette for Saccharomyces cerevisiae.FEMS yeast research · 2013Article
- A novel, lactase-based selection and strain improvement strategy for recombinant protein expression in Kluyveromyces lactis.Microbial cell factories · 2012Article
- A new method for repeated "self-cloning" promoter replacement in Saccharomyces cerevisiae.Molecular biotechnology · 2011Article
- A set of aspartyl protease-deficient strains for improved expression of heterologous proteins in Kluyveromyces lactis.FEMS yeast research · 2011Article
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4 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
The yeast Kluyveromyces lactis has been extensively used as a host for heterologous protein expression. A necessary step in the construction of a stable expression strain is the introduction of an integrative expression vector into K. lactis cells, followed by selection of transformed strains using either medium containing antibiotic (e.g., G418) or nitrogen-free medium containing acetamide. In this study, we show that selection using acetamide yields K. lactis transformant populations nearly completely comprised of strains bearing multiple tandem insertions of the expression vector pKLAC1 at the LAC4 chromosomal locus, whereas an average of 16% of G418-selected transformants are multiply integrated. Additionally, the average copy number within transformant populations doubled when acetamide was used for selection compared to G418. Finally, we demonstrate that the high frequency of multicopy integration associated with using acetamide selection can be exploited to rapidly construct expression strains that simultaneously produce multiple heterologous proteins or multisubunit proteins, such as Fab antibodies.
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