AvrSr35 诱导的 Sr35 抗性体的组装和纯化及冷冻电镜结构测定。
Assembly and purification of AvrSr35-induced Sr35 resistosome and determination of its structure by cryo-EM.
机构信息
Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, the Key Laboratory of Innate Immune Biology of Fujian Province, Biomedical Research Center of South China, Key Laboratory of OptoElectronic Science and Technology for Medicine of the Ministry of Education, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China.
Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, the Key Laboratory of Innate Immune Biology of Fujian Province, Biomedical Research Center of South China, Key Laboratory of OptoElectronic Science and Technology for Medicine of the Ministry of Education, College of Life Sciences, Fujian Normal University, Fuzhou 350117, China.
出版信息
STAR Protoc. 2022 Oct 31;3(4):101796. doi: 10.1016/j.xpro.2022.101796. eCollection 2022 Dec 16.
Sr35, a coiled-coil nucleotide-binding leucine-rich repeat receptor (CC-NLR) from the wheat species can directly recognize the pathogen avirulence factor AvrSr35 and confers immunity against wheat stem rust caused by f.sp. race Ug99. Assembly of a stable Sr35 resistosome induced by AvrSr35 is usually limited by protein expression and low assembly efficiency. Here, we describe the expression and purification of AvrSr35 and Sr35, assembly of Sr35 resistosome for structure determination by cryo-EM. For complete details on the use and execution of this protocol, please refer to Zhao et al. (2022).
Sr35 是一种来自小麦属的卷曲螺旋核苷酸结合亮氨酸丰富重复受体 (CC-NLR)，可直接识别病原菌无毒因子 AvrSr35，并赋予其对由 f.sp. 引起的小麦茎锈病的免疫性。Ug99 。由 AvrSr35 诱导的稳定 Sr35 抗性体的组装通常受到蛋白表达和低组装效率的限制。在这里，我们描述了 AvrSr35 和 Sr35 的表达和纯化，以及 Sr35 抗性体的组装，用于通过 cryo-EM 进行结构测定。有关此协议的使用和执行的完整详细信息，请参见 Zhao 等人。（2022 年）。
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