Stable, selectable, integrative DNA transformation in Physarum
1993
Burland, T.G. | Bailey, J. | Pallotta, D. | Dove, W.F.
The Physarum polycephalum actin promoter, PardC, can drive transient expression of heterologous genes in Physarum amoebae. The hph gene, encoding hygromycin (Hy) phosphotransferase, can confer resistance to Hy on a broad spectrum of organisms. When PardC is translationally fused to hph and transformed into yeasts on high-copy-number vectors, the yeasts become Hy resistant (Hy(R)), showing that PardC-hph is a functional, selectable genetic element. To establish a stable transformation system for Physarum, we electroporated plasmids bearing PardC-hph into Physarum amoebae and then selected for Hy(R) transformants. We show that Hy(R) amoebae arise upon the stable integration of PardC-hph into the nuclear genome in single copy. These results establish a transformation system that can be used to add plasmid-borne genetic information to Physarum.
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