The URA1 gene, encoding dihydroorotate dehydrogenase of the pyrimidine pathway, cloned into pUC18 (pUral-1) was used to develop an homologous transformation system for the cultivated mushroom Agrocybe aegerita. Protoplasts of a ural auxotrophic strain were transformed by electroporation with efficie
Transformation ofSchizosaccharomyces pombeby non-homologous, unstable integration of plasmids in the genome
✍ Scribed by A. P. H. Wright; K. Maundrell; S. Shall
- Book ID
- 104778249
- Publisher
- Springer-Verlag
- Year
- 1986
- Tongue
- English
- Weight
- 786 KB
- Volume
- 10
- Category
- Article
- ISSN
- 0172-8083
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✦ Synopsis
In the fission yeast, Schizosaccharomyces pombe, transformation with recombinant plasmids always results in a high proportion of mitotically unstable transformants. This suggested that specialised (ARS) sequences might not be required for autonomous replication of plasmids in S. pombe, contrary to the situation in Saccharomyces cerevisiae. We have shown that specialised ARS sequences, analogous to those in S. cerevisiae, do exist in S. pombe, supporting the view that ARS elements are a general feature of eukaryotes. In addition, there is a further mechanism of plasmid maintenance which involves homologous and non-homologous integration into, and excision from the genome.
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