| Heavy Metal Stress Assay of Caenorhabditis elegans
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Author:
Date:
2017-06-05
[Abstract] Organisms have developed many protective systems to reduce the toxicity from heavy metals. The nematode Caenorhabditis elegans has been widely used to determine the protective mechanisms against heavy metals. Responses against heavy metals can be monitored by expression of reporter genes, while sensitivity can be determined by quantifying growth or survival rate following exposure to heavy metals.
[摘要] 生物开发了许多保护系统，以减少重金属的毒性。线虫秀丽隐杆线虫已广泛用于确定重金属的保护机制。可以通过报告基因的表达来监测对重金属的反应，而敏感性可以通过量化暴露于重金属后的生长或存活率来确定。
背景 一些重金属如砷，镉和汞已知对包括人类在内的大多数生物有害（Valko et al。，2005）。为了降低这些金属的毒性，生物开发出各种保护系统。线虫秀丽隐杆线虫已被用于了解重金属的保护机制。以前的研究表明，许多基因，如解毒酶，转录因子和信号传导因子都参与了该生物体中重金属的保护（Broeks et al。，1996; Mizuno et al。 ，2004; Inoue等人，2005; Schwartz等人，2010）。除了测量报告基因表达外，生存力和生长的测定通常用于监测重金属在C中的影响。线虫。在本协议中，我们描述了使用C测定砷，铜和镉的方法。线虫。
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| Purification of Flagellin from Acidovorax avenae and Analysis of Plant Immune Responses Induced by the Purified Flagellin
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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。
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| Total RNA Extraction from Grape Berry Skin for Quantitative Reverse Transcription PCR and Microarray Analysis
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Author:
Date:
2016-04-05
[Abstract] Extraction of high quality RNA is an essential step for quantitative reverse transcription PCR (qRT-PCR) and microarray analysis. However, it is not easy to extract high quality RNA from fruit materials, which contain high amounts of polysaccharides, lipids and secondary metabolites. Wan and Wilkins (1994) had developed ‘Hot Borate Method’ to isolate high quality RNA. Here, we describe a modified protocol of the ‘Hot Borate Method’ to isolate high quality RNA from grape berry skin for qRT-PCR and microarray analysis (Suzuki et al., 2015a; Suzuki et al., 2015b).
[摘要] 高质量RNA的提取是定量逆转录PCR（qRT-PCR）和微阵列分析的必要步骤。 然而，从含有大量多糖，脂质和次级代谢物的水果材料中提取高质量RNA是不容易的。 Wan和Wilkins（1994）开发了"热硼酸盐方法"来分离高质量的RNA。 在这里，我们描述了"热硼酸盐方法"的修饰的协议，从葡萄浆果皮肤中分离高质量的RNA用于qRT-PCR和微阵列分析（Suzuki等人，2015a; Suzuki等人 al。，2015b）。
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