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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.
P K Howard, and
K G Ahern, and
R A Firtel
March 2009,
Plasmid,
P K Howard, and
K G Ahern, and
R A Firtel
December 2008,
FASEB journal : official publication of the Federation of American Societies for Experimental Biology,
P K Howard, and
K G Ahern, and
R A Firtel
December 1982,
Proceedings of the National Academy of Sciences of the United States of America,
P K Howard, and
K G Ahern, and
R A Firtel
January 2022,
Cells,
P K Howard, and
K G Ahern, and
R A Firtel
January 2019,
The International journal of developmental biology,
P K Howard, and
K G Ahern, and
R A Firtel
May 1988,
Gene,
P K Howard, and
K G Ahern, and
R A Firtel
January 1975,
Basic life sciences,
P K Howard, and
K G Ahern, and
R A Firtel
December 2000,
The International journal of developmental biology,
P K Howard, and
K G Ahern, and
R A Firtel
January 1999,
Biology of the cell,
P K Howard, and
K G Ahern, and
R A Firtel
October 1994,
Molecular and cellular biochemistry,
