Welsh Lydia R J, Whisson Stephen C
Cell and Molecular Sciences Department, The James Hutton Institute, Dundee, UK.
Methods Mol Biol. 2025;2892:35-47. doi: 10.1007/978-1-0716-4330-3_3.
At the core of assays to understand the role(s) of specific genes is the ability to stably transfer genes into Phytophthora through transformation. A key method for achieving this has been based on polyethylene glycol (PEG)/CaCl transformation of protoplasts, but efficiency has often been low. Improving transformation efficiency is necessary for many applications, such as gene knockouts. Here we describe improvements through successive rounds of "mock" transformation, leading to improved efficiency in Phytophthora infestans and other species.
理解特定基因作用的检测方法的核心是通过转化将基因稳定转移到疫霉中的能力。实现这一目标的关键方法是基于原生质体的聚乙二醇（PEG）/氯化钙转化，但效率往往较低。提高转化效率对于许多应用（如基因敲除）来说是必要的。在这里，我们描述了通过连续几轮“模拟”转化所取得的改进，从而提高了致病疫霉和其他物种的转化效率。
