ArticleCurrent genetics1992
Multiple-copy integration of the alpha-galactosidase gene from Cyamopsis tetragonoloba into the ribosomal DNA of Kluyveromyces lactis.
Article in Current genetics, 1992. 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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Who cites it
15 citing papers in PubMed, 57 citations in OpenAlex.
- Comparison of Yeasts as Hosts for Recombinant Protein Production.Microorganisms · 2018Review
- Heterologous expression of glucose oxidase in the yeast Kluyveromyces marxianus.Microbial cell factories · 2010Article
- Kluyveromyces lactis LAC4 promoter variants that lack function in bacteria but retain full function in K. lactis.Applied and environmental microbiology · 2005Article
- A wide-range integrative yeast expression vector system based on Arxula adeninivorans-derived elements.Journal of industrial microbiology & biotechnology · 2004Article
- Autocloning and amplification of LIP2 in Yarrowia lipolytica.Applied and environmental microbiology · 2000Article
- Use of the KlADH4 promoter for ethanol-dependent production of recombinant human serum albumin in Kluyveromyces lactis.Applied and environmental microbiology · 1999Article
- Heterologous protein secretion directed by a repressible acid phosphatase system of Kluyveromyces lactis: characterization of upstream region-activating sequences in the KIPHO5 gene.Applied and environmental microbiology · 1998Article
- Expression and secretion of a thermostable bacterial xylanase in Kluyveromyces lactis.Applied and environmental microbiology · 1997Article
- Glucoamylase gene fusions alleviate limitations for protein production in Aspergillus awamori at the transcriptional and (post) translational levels.Applied and environmental microbiology · 1997Article
- Analysis of heterologous protein production in defined recombinant Aspergillus awamori strains.Applied and environmental microbiology · 1996Article
- Expression of the Arxula adeninivorans glucoamylase gene in Kluyveromyces lactis.Applied microbiology and biotechnology · 1996Article
- Physiological and technological aspects of large-scale heterologous-protein production with yeasts.Antonie van Leeuwenhoek · 1995Review
- Multiple-copy integration in the yeast Yarrowia lipolytica.Current genetics · 1994Article
- Expression of an alpha-galactosidase gene under control of the homologous inulinase promoter in Kluyveromyces marxianus.Applied microbiology and biotechnology · 1993Article
- The yeast Kluyveromyces lactis as an efficient host for heterologous gene expression.Antonie van LeeuwenhoekReview
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We have developed a vector system for high-copy-number integration into the ribosomal DNA of the yeast Kluyveromyces lactis. This system is analogous to the pMIRY-system developed for Saccharomyces cerevisiae. Plasmids containing a portion of K. lactis rDNA for targeted homologous recombination, as well as the S. cerevisiae TRP1 gene with various promoter deletions, were constructed and, after transformation to K. lactis, analyzed for both copy number and stability. These plasmids were found to be present in about 60 copies per cell and were stably maintained during growth under non-selective conditions. Using this vector system, we expressed a fusion construct containing the S. cerevisiae GAL7 promoter, the SUC2 (invertase) signal sequence and the gene coding for alpha-galactosidase from the plant Cyamopsis tetragonoloba. Although the maximum copy number of these integrated plasmids was only about 15, we nevertheless obtained a high level of alpha-galactosidase production (250 mg/l) with a secretion efficiency of about 95%. When compared to extrachromosomal K. lactis vectors containing the same fusion construct, the multicopy integrants showed a much higher alpha-galactosidase production level and a considerably higher stability under non-selective conditions.
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