Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1988 Mar 25;16(6):2613-23.
doi: 10.1093/nar/16.6.2613.

Establishment of a transient expression system for Dictyostelium discoideum

P K Howard  1 , K G Ahern, R A Firtel
Affiliations
Free PMC article

Establishment of a transient expression system for Dictyostelium discoideum

P K Howard et al. Nucleic Acids Res. .
Free PMC article

Abstract

We have established a rapid and sensitive transient expression system for Dictyostelium discoideum. We constructed a gene fusion containing the promoter from the Dictyostelium Actin 15 gene fused to the firefly luciferase gene. The enzymatic activity of this gene fusion, expressed at very high levels in stable transformants, was measured to determine optimum conditions for transient expression using electroporation to introduce the DNA into cells. With these conditions, we show that a luciferase gene fusion driven by a prestalk, cell-type specific promoter from the pst-cathepsin gene expresses luciferase at the appropriate developmental stage. In addition, we present results suggesting that the system will be useful for expressing genes in non-axenic cell lines. Finally, we observe that electroporation is more efficient for obtaining stable transformations than the standard calcium phosphate procedure using extrachromosomally replicating shuttle vectors but less efficient for vectors that integrate into the Dictyostelium chromosomes.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Mol Cell Biol. 1984 Dec;4(12):2890-8 - PubMed
    1. Gene. 1987;59(1):99-106 - PubMed
    1. Cell. 1982 Apr;28(4):793-800 - PubMed
    1. EMBO J. 1987 Dec 20;6(13):4143-8 - PubMed
    1. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7161-5 - PubMed

Publication types