Desalted deep sea water increases transformation and homologous recombination efficiencies in Dictyostelium discoideum.
The life cycle of Dictyostelium discoideum consists of many cellular and developmental aspects. By virtue of its relatively high transformation efficiency and a small haploid genome, this organism has proven to be advantageous for characterizing gene functions. However, a much higher transformation efficiency is required as one of the prerequisites for unraveling gene function on a genome-wide scale. In this study, we describe the positive effect of desalted deep sea water, when used as a solvent medium, on the transformation and homologous recombination efficiencies in Dictyostelium. A standard Dictyostelium medium HL5 containing desalted deep sea water, HL5dsw, distinctly increased both the transformation and homologous recombination efficiencies by approximately 2- to 3-fold. Furthermore, we observed that the growth of cells in HL5dsw both before and after electroporation contributed to the increase in transformation efficiency. These results indicate that a simple modification of the solvent medium remarkably enhanced the isolation of transformants and gene-targeted clones, which had previously been difficult to isolate.
Kuwayama
Hidekazu
H
Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan. hidekuwayama@biol.tsukuba.ac.jp
Nagasaki
Akira
A
D000818:Animals; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D004023:Dictyostelium / Q000235:genetics* / Q000302:isolation & purification; D018274:Electroporation; D011995:Recombination, Genetic / Q000187:drug effects*; D012623:Seawater / Q000737:chemistry*; D014170:Transformation, Genetic / Q000187:drug effects*
D003470:Culture Media