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Stable, selectable, integrative DNA transformation in Physarum

T G Burland1, J Bailey, D Pallotta

  • 1McArdle Laboratory for Cancer Research, Madison, WI 53706.

Gene
|October 15, 1993
PubMed
Summary
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Researchers developed a stable transformation system for Physarum polycephalum using the PardC promoter and hygromycin resistance gene. This system enables the stable integration of genetic elements into the Physarum genome.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • The Physarum polycephalum actin promoter (PardC) drives transient gene expression in amoebae.
  • The hygromycin (Hy) phosphotransferase (hph) gene confers resistance to hygromycin in various organisms.
  • A functional, selectable genetic element (PardC-hph) was demonstrated in yeast.

Purpose of the Study:

  • To establish a stable transformation system for Physarum polycephalum.
  • To enable the stable integration of heterologous genes into the Physarum genome.

Main Methods:

  • Translational fusion of PardC with the hph gene.
  • Transformation of yeast with high-copy-number vectors containing PardC-hph.
  • Electroporation of Physarum amoebae with plasmids carrying PardC-hph.

Related Experiment Videos

  • Selection of hygromycin-resistant (HyR) transformants.

    • Main Results:

    • PardC-hph fusion conferred hygromycin resistance in yeast.
    • Stable integration of PardC-hph into the Physarum nuclear genome was achieved.
    • HyR transformants arose from single-copy integration of PardC-hph.
    • A functional, selectable genetic element was successfully introduced into Physarum.

    Conclusions:

    • A stable transformation system for Physarum polycephalum has been established.
    • This system allows for the addition of plasmid-borne genetic information to Physarum.
    • The PardC-hph construct serves as a selectable marker for stable Physarum transformation.