title: Stable, selectable, integrative DNA transformation in Physarum.
abstract: 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 (HyR), 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 HyR transformants. We show that HyR 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.
authors.affiliation: McArdle Laboratory for Cancer Research, Madison, WI 53706.
authors.forename: T G
authors.initials: TG
authors.lastname: Burland
authors.forename: J
authors.initials: J
authors.lastname: Bailey
authors.forename: D
authors.initials: D
authors.lastname: Pallotta
authors.forename: W F
authors.initials: WF
authors.lastname: Dove
mesh_terms: D000199:Actins / Q000235:genetics; D000818:Animals; D015139:Blotting, Southern; D002934:Cinnamates; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000378:metabolism; D010641:Phenotype; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D010804:Physarum / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014170:Transformation, Genetic
chemical_list: D000199:Actins; D002934:Cinnamates; D006921:Hygromycin B; C026273:hygromycin A; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase