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Nuclear plasmids in the Dictyostelium slime molds
J E Hughes1, H Ashtorab, D L Welker
1Dept. of Biology, Utah State University, Logan 84322-5500.
Developmental Genetics
|January 1, 1988
Summary
Cellular slime molds possess unique circular nuclear plasmids. Researchers developed efficient methods to introduce these Dictyostelium plasmids into cells, enabling foreign DNA delivery.
Area of Science:
- Molecular Biology
- Genetics
- Eukaryotic Cell Biology
Background:
- Cellular slime molds are rare eukaryotes with nuclear plasmids.
- Dictyostelium species harbor diverse, nonhomologous circular DNA plasmids.
- Plasmids are found in approximately 20% of Dictyostelium isolates and organized in nucleosomes.
Purpose of the Study:
- To investigate the cotransformation of endogenous Dictyostelium plasmids into D. discoideum.
- To develop efficient extrachromosomal vectors for genetic manipulation in D. discoideum.
- To assess the stability and utility of recombinant plasmids for introducing foreign DNA.
Main Methods:
- Cotransformation of Dictyostelium discoideum using the G418 resistance shuttle vector B10S.
- Construction of recombinant plasmids based on the D. discoideum plasmid Ddp2 and a resistance gene.
- Analysis of transformant recovery frequency, plasmid copy number, and stability.
Main Results:
- Cotransformation of endogenous D. discoideum plasmids yielded higher frequencies than plasmids from other species.
- Recombinant plasmids based on Ddp2 achieved transformation frequencies up to 10^-4 per input cell.
- Constructed vectors demonstrated stable maintenance at high copy number without selection.
Conclusions:
- Endogenous Dictyostelium plasmids can be efficiently cotransformed into D. discoideum.
- Recombinant plasmids derived from Dictyostelium plasmids serve as effective extrachromosomal vectors.
- These vectors facilitate the stable introduction of foreign DNA into D. discoideum.