| Quantification of Ethylene Production in Tomato Leaves Infected by Xanthomonas euvesicatoria
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Author:
Date:
2016-02-05
[Abstract] Ethylene is a gaseous plant hormone controlling fruit ripening, flower opening, leaf senescence as well as abscission, and disease symptom development. Ethylene plays a critical role in the bacterial pathogen Xanthomonas euvesicatoria (X. euvesicatoria)-elicited symptom development in tomato. This protocol describes the measurement of ethylene gas produced by tomato leaves infected with X. euvesicatoria. Infected leaflets are placed in a glass tube for 30 min without sealing. The glass tubes are then capped with a septa stopper, and incubated for an hour. A 1 ml gas sample is removed from the tube using a syringe and then injected into a gas chromatograph to quantify ethylene gas levels. This protocol will be applicable for other plants with other pathogens with ...
[摘要] 乙烯是控制果实成熟，开花，叶衰老以及脱落和疾病症状发展的气态植物激素。 乙烯在细菌病原体黄单胞菌属（Xanthomonas euvesicatoria）（ X。euvesicatoria ）中起到关键作用 - 引起番茄中的症状发展。 该方案描述了用感染了X的番茄叶产生的乙烯气体的测量。 euvesicatoria 。 将感染的小叶放置在玻璃管中30分钟而不密封。 然后将玻璃管用隔膜塞子盖上，并孵育1小时。 使用注射器从管中移出1ml气体样品，然后注入气相色谱仪中以定量乙烯气体水平。 该协议将适用于其他具有修改的其他病原体的植物。
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| CytoTrap Two-Hybrid Screening Assay
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Author:
Date:
2014-12-05
[Abstract] CytoTrap two-hybrid system provides an alternate strategy to detect protein-protein interactions in yeast. In this system, bait protein is fused with human son of sevenless (hSos) protein (Li et al., 1993), and a cDNA library or prey protein is expressed by fusion with myristoylation signal which anchors the prey fusion protein to yeast cell membrane. Protein interaction between bait and prey proteins recruits the hSos protein to the cell membrane, where hSos activates the Ras signaling pathway, leading to the survival of temperature-sensitive Saccharomyces cerevisiae (S. cerevisiae) strain cdc25H at 36 °C. In the CytoTrap two-hybrid system, detection of protein interaction occurs in the cytoplasm near cell membrane and is not dependent on transcription ...
[摘要] CytoTrap双杂交系统提供了检测酵母中蛋白质 - 蛋白质相互作用的替代策略。在该系统中，诱饵蛋白与七（hSos）蛋白质的人子融合（Li等，1993），cDNA文库或猎物蛋白质与肉豆蔻酰化信号的融合表达，将猎物融合蛋白锚定到酵母细胞膜。诱饵和猎物蛋白之间的蛋白质相互作用将hSos蛋白质募集到细胞膜上，其中hSos激活Ras信号通路，导致温度敏感的酿酒酵母（S.cerevisiae）菌株cdc25H在36℃下的存活。在CytoTrap双杂交系统中，蛋白质相互作用的检测发生在细胞膜附近的细胞质中，不依赖于报告基因的转录激活。因此，该系统对于识别需要在细胞质中进行翻译后修饰的转录因子和蛋白质的相互作用伴侣特别有用，其在常规基于反式激活的酵母双杂交系统中不能用作诱饵蛋白。在这里，我们描述了来自模拟植物拟南芥的cDNA文库的构建以及用于筛选来自该文库的AtSR1 / CAMTA3，Ca2 + / CaM调节的转录因子的相互作用蛋白的程序。该过程可以适应于识别来自其他生物体的感兴趣的蛋白质的相互作用的伴侣。
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