Edskes Herman K, Kryndushkin Dmitry, Shewmaker Frank, Wickner Reed B
Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-0830.
Department of Pharmacology, Uniformed Services University for the Health Sciences, Bethesda, Maryland 20814.
Cold Spring Harb Protoc. 2017 Feb 1;2017(2):2017/2/pdb.prot089037. doi: 10.1101/pdb.prot089037.
Transfection of yeast with amyloid filaments, made from recombinant protein or prepared from extracts of cells infected with a prion, has become an important method in characterizing yeast prions. Here, we describe a method for transmission of [URE3] with Ure2p amyloid that is based on a previously published protocol for transfection with Sup35p filaments to make cells [PSI+]. This method may be used for other prions by changing just the amyloid source, host strain, and plating medium.
用由重组蛋白制成或从感染朊病毒的细胞提取物制备的淀粉样细丝转染酵母，已成为表征酵母朊病毒的一种重要方法。在此，我们描述了一种用Ure2p淀粉样蛋白传播[URE3]的方法，该方法基于先前发表的用Sup35p细丝转染以使细胞成为[PSI+]的方案。通过仅改变淀粉样蛋白来源、宿主菌株和平板培养基，该方法可用于其他朊病毒。
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