| Analysis of Autophagic Activity Using ATG8 Lipidation Assay in Arabidopsis thaliana
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Author:
Date:
2018-06-20
[Abstract] As a fundamental metabolic pathway to degrade and recycle cellular cargos, autophagy is highly induced upon stress, starvation and senescence conditions in plants. A double-membrane structure named autophagosome will form during this process for cargo sequestration and delivery into the vacuole.
A number of regulators have been characterized in plants, including the autophagy-related (ATG) proteins and other plant-specific proteins. Among them, ATG8 will undergo a lipidation process to become a membrane-bound ATG8-phosphatidylethanolamine form and mark the growing autophagosomal ...
[摘要] 作为降解和回收细胞货物的基本代谢途径，自噬在植物的胁迫，饥饿和衰老条件下被高度诱导。 在这个过程中，将形成一个称为自噬体的双膜结构，用于货物隔离和输送到液泡中。
已经在植物中表征了许多调控因子，包括自噬相关（ATG）蛋白和其他植物特异性蛋白。 其中，ATG8将经历脂化过程以成为膜结合的ATG8-磷脂酰乙醇胺形式并标记日益增长的自噬体膜以及完成的自噬体。 因此，ATG8被认为是自噬体的标志; 并且膜结合形式的ATG8的生物化学检测被用作测量自噬活性的主要方法之一。 在这里，我们描述了使用拟南芥幼苗检测ATG8-PE形式的ATG8脂化测定法。
【背景】自噬是调节受损细胞器的大量降解和不需要的细胞内容物的基本代谢过程。在自噬过程中，称为自噬体的双膜结构将形成并将货物递送到液泡中以降解。自噬相关蛋白（ATG）需要调节自噬活性（Liu and ...
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| Isolation of Radiolabeled Poliovirus Particles from H1 HeLa Cells
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Author:
Date:
2013-08-20
[Abstract] The following protocol describes the isolation of radioactive viral and subviral particles from infected cells. This protocol has been written for isolation of poliovirus particles from H1 HeLa cells. Infection protocol and timing of [35S] methionine labeling and particle collection should be tailored to the virus of interest. Following isolation of the viral particles, the viral proteins present in these particles may be separated by gel electrophoresis and visualized by autoradiography.
[摘要] 以下方案描述了从感染的细胞中分离放射性病毒和亚病毒颗粒。 该协议是为了从H1 HeLa细胞分离脊髓灰质炎病毒颗粒而编写的。 [35]甲硫氨酸标记和颗粒收集的感染方案和时机应该针对感兴趣的病毒定制。 分离病毒颗粒后，可以通过凝胶电泳分离存在于这些颗粒中的病毒蛋白，并通过放射自显影进行显现。
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