RRC ID 36699
Author Kuwayama H, Nagasaki A.
Title Desalted deep sea water increases transformation and homologous recombination efficiencies in Dictyostelium discoideum.
Journal J Mol Microbiol Biotechnol
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.
Volume 14(4)
Pages 157-62
Published 2008-1-1
DOI 10.1159/000107371
PII 000107371
PMID 17693704
MeSH Animals Culture Media / chemistry Culture Media / pharmacology* Dictyostelium / genetics* Dictyostelium / isolation & purification Electroporation Recombination, Genetic / drug effects* Seawater / chemistry* Transformation, Genetic / drug effects*
IF 0.803
Times Cited 10
WOS Category BIOTECHNOLOGY & APPLIED MICROBIOLOGY MICROBIOLOGY
Resource
Cellular slime molds