| Purification of Flagellin from Acidovorax avenae and Analysis of Plant Immune Responses Induced by the Purified Flagellin
|
|
Author:
Date:
2016-08-20
[Abstract] Plants sense potential pathogens by recognizing conserved pathogen-associated molecular patterns (PAMPs) that cause PAMP-triggered immunity (PTI) including the generation of reactive oxygen species, callose deposition, and expression of several PTI-related genes. Acidovorax avenae is a Gam-negative bacterium that causes a seedling disease characterized by the deposition of brown stripes on the sheaths of infected plants. We previously reported that flagellin isolated from the rice avirulent A. avenae N1141 strain induces PTI, while flagellin isolated from the rice virulent A. avenae K1 strain does not induce PTI. To examine the molecular mechanism of specific PTI induction by N1141 flagellin, highly purified flagellin from N1141 or K1 strains is required. Here, ...
[摘要] 植物通过识别引起PAMP触发的免疫（PTI）的保守的病原体相关分子模式（PAMP）来感测潜在的病原体，包括活性氧的产生，胼lose质沉积和几个PTI相关基因的表达。 酸性弧菌是一种革兰氏阴性细菌，其导致幼苗疾病，其特征在于在感染的植物的鞘上沉积棕色条纹。 我们以前报告了从无毒力的大米中分离的鞭毛蛋白。 avenae N1141菌株诱导PTI，而从水稻毒力分离的鞭毛蛋白。 avenae K1株不诱导PTI。 为了检查N1141鞭毛蛋白的特异性PTI诱导的分子机制，需要来自N1141或K1菌株的高度纯化的鞭毛蛋白。 在这里，我们描述了一种高质量的纯化方法。 avenae鞭毛蛋白，并用于PTI诱导研究。
|
|
|
| Colon Tissue Immunoelectron Microscopy
|
|
Author:
Date:
2013-10-20
[Abstract] The method described here is intended to study intracellular localization of proteins in colon cells. This protocol was used to localize IRE1β in the endoplasmic reticulum membrane. We used anti-IRE1β antibody raised in guinea pig for this purpose. We also studied the location of BiP (also known as GRP78), with the antibody raised in rabbit. Both antibodies used with appropriate gold particle-conjugated secondary antibodies gave good results. Primary mouse antibodies are not recommended because secondary anti-mouse antibodies also react with the internal mouse epitopes.
[摘要] 本文所述的方法旨在研究结肠细胞中蛋白质的细胞内定位。 该方案用于定位IRE1β在内质网膜中。 我们使用在豚鼠中产生的抗IRE1β抗体用于此目的。 我们还研究了在兔中产生的抗体的BiP（也称为GRP78）的位置。 与合适的金颗粒缀合的二抗一起使用的两种抗体都给出了良好的结果。 不推荐初级小鼠抗体，因为次级抗小鼠抗体也与内部小鼠表位反应。
|
|
|
| Transmission Electron Microscopy (TEM) Protocol: Observation Details within Cells
|
|
Author:
Date:
2013-07-05
[Abstract] Transmission electron microscopy is a technique for observing the fine details of organelles in cells or tissues. This protocol is to be used to exam the membrane structure in cells with or without virus infection. Modifications should be made if users want to get images from tissues.
[摘要] 透射电子显微镜是用于观察细胞或组织中细胞器的细节的技术。 该协议用于检查有或没有病毒感染的细胞中的膜结构。 如果用户想要从组织获得图像，应该进行修改。
|
|
|