Evidence for autonomous replication and stabilization of recombinant plasmids in the transformants of yeastHansenula polymorpha
✍ Scribed by L. P. Tikhomirova; R. N. Ikonomova; E. N. Kuznetsova
- Publisher
- Springer-Verlag
- Year
- 1986
- Tongue
- English
- Weight
- 711 KB
- Volume
- 10
- Category
- Article
- ISSN
- 0172-8083
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✦ Synopsis
Moscow region, 142292, USSR Summary. For the transformation of the yeast Hansenula polymorpha we have constructed a set of hybrid plasmids carrying the LEU2 gene ofSaccharomyces cerevisiae as a selective marker and fragments of mitochondrial DNA of Candida utilis and H. polymorpha or chromosomal DNA fragments of H. polymorpha as replicator sequences. The replication properties of chimeric plasmids in the yeast H. polymorpha were investigated. We showed that for plasmids propagated autonomously in this yeast the plasmid monomers could be detected in the transformants only during the immediate time after the transformation event. Further growth under selective conditions led to the selection of polymeric forms of plasmid DNA as it was clearly shown for transformants carrying cosmid pL2 with mtDNA fragment of C utilis. Such transformants carrying polymerized plasmids showed a remarkably increased stability of the transformed phenotype. Cosmid pL2 was able to shuttle between Escherichia coli, S. cerevisiae and H. polymorpha, whereas plasmids with DNA fragments from H. polymorpha did not transform S. cerevisiae effectively.