Amplification of purified prions in vitro.
Amplification of purified prions in vitro.
复制标题
纯化朊病毒的体外扩增。
DOI：
10.1007/978-1-59745-234-2_9
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Rees,JudyR
中科院分区：
文献类型：
--
作者：
Supattapone,Surachai;Deleault,NathanR;Rees,JudyR
The infectious agents of prion diseases are unorthodox, and they seem to be composed primarily of a misfolded glycoprotein called the prion protein (PrP). Replication of prion infectivity is associated with the conversion of PrP from its normal, cellular form (PrPC) into a pathogenic form (PrPSc), which is characterized biochemically by relative detergent insolubility and protease resistance. Several techniques have been developed in which PrPCmolecules can be converted into the PrPScconformation in vitro (1–8). These biochemical techniques recapitulate several specific aspects of in vivo prion propagation (1–3), and one method, the protein misfolding cyclic amplification technique, also has been shown to amplify infectivity (5). In this chapter, we describe a method for amplifying PrPScmolecules from hamster prions in vitro using purified substrates. Specific protocols for substrate preparation, reaction mixture, and product detection are explained. Purified PrPScamplification assays are currently being used to study the biochemical mechanism of prion formation.
