Hosoe Naoi, Keima Takuya, Fujimoto Yuji, Hagiwara-Komoda Yuka, Hashimoto Masayoshi, Maejima Kensaku, Namba Shigetou, Yamaji Yasuyuki
Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
Methods Mol Biol. 2019;2028:145-151. doi: 10.1007/978-1-4939-9635-3_8.
Plant viruses use numerous host factors for efficient replication of the viral genome. Protoplasts, plant cells from which cell walls are removed, are the useful system to analyze the virus translation and replication in vivo. Here, we report a protocol for preparation of protoplasts from Arabidopsis thaliana leaves and transfection of plasmids to the protoplasts. Protoplasts isolated from the loss-of-function mutant of viral host factor(s) would be helpful to analyze the function of host factors in virus infection cycles.
植物病毒利用多种宿主因子来实现病毒基因组的高效复制。原生质体是去除细胞壁后的植物细胞，是分析病毒在体内翻译和复制的有用系统。在此，我们报告一种从拟南芥叶片制备原生质体并将质粒转染至原生质体的方法。从病毒宿主因子功能缺失突变体中分离出的原生质体，将有助于分析宿主因子在病毒感染周期中的功能。
