Alibaud Laeticia, Cosson Pierre, Benghezal Mohammed
Université de Genève, Athelas SA, Geneva, Switzerland.
Biotechniques. 2003 Jul;35(1):78-80, 82-3. doi: 10.2144/03351st03.
Dictyostelium discoideum has been used as a genetically tractable model organism to study many biological phenomena. High-efficiency transformation is a prerequisite for successful genetic screens such as mutant complementation, identification of suppressor genes, or insertional mutagenesis. Although exponential decay electroporation is the standard transformation technique for D. discoideum, its efficiency is relatively low and its reproducibility is weak. Here we optimized the oscillating electroporation technique for D. discoideum transformation and compared it to the exponential decay electroporation. A 20-fold increase in the efficiency was resproducibly achieved. This alternative electroporation technique should facilitate future genetic approaches in D. discoideum.
盘基网柄菌已被用作一种遗传上易于操作的模式生物来研究许多生物学现象。高效转化是成功进行基因筛选（如突变体互补、抑制基因鉴定或插入诱变）的先决条件。尽管指数衰减电穿孔是盘基网柄菌的标准转化技术，但其效率相对较低且重复性较差。在此，我们优化了用于盘基网柄菌转化的振荡电穿孔技术，并将其与指数衰减电穿孔进行了比较。可重复地实现了效率提高20倍。这种替代的电穿孔技术应有助于盘基网柄菌未来的遗传学研究方法。
