title: Desalted deep sea water increases transformation and homologous recombination efficiencies in Dictyostelium discoideum.
abstract: 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.
authors: Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki, Japan. hidekuwayama@biol.tsukuba.ac.jp
authors: Hidekazu
authors: H
authors: Kuwayama
authors: Akira
authors: A
authors: Nagasaki
mesh_terms: 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*
chemical_list: D003470:Culture Media