蛋白质错误折叠循环扩增方法
Methods of Protein Misfolding Cyclic Amplification.
作者信息
Makarava Natallia, Savtchenko Regina, Baskakov Ilia V
机构信息
Department of Anatomy and Neurobiology, Center for Biomedical Engineering and Technology, University of Maryland School of Medicine, 111 Penn St., Baltimore, MD, 21201, USA.
出版信息
Methods Mol Biol. 2017;1658:169-183. doi: 10.1007/978-1-4939-7244-9_13.
Protein misfolding cyclic amplification (PMCA) amplifies infectious prions in vitro. Over the past decade, PMCA has become an essential tool in prion research. The current chapter describes in detail the PMCA format with beads (PMCAb) and several methods that rely on PMCAb for assessing strain-specific prion amplification rates, for selective amplification of subtypes of PrP from a mixture, and a PMCAb approach that can replace animal titration of scrapie material. Development of PMCAb-based methodology is important for addressing a number of research topics including prion strain evolution, selection and adaptation, strain-typing, prion detection, and biochemical requirements for prion replication.
蛋白质错误折叠循环扩增（PMCA）可在体外扩增传染性朊病毒。在过去十年中，PMCA已成为朊病毒研究中的一项重要工具。本章详细描述了带珠粒的PMCA形式（PMCAb）以及几种依赖PMCAb来评估毒株特异性朊病毒扩增率、从混合物中选择性扩增PrP亚型的方法，还有一种可替代对羊瘙痒病材料进行动物滴定的PMCAb方法。基于PMCAb的方法学发展对于解决许多研究课题至关重要，这些课题包括朊病毒株的进化、选择与适应、毒株分型、朊病毒检测以及朊病毒复制的生化要求。
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