| Detection of Reactive Oxygen Species in Oryza sativa L. (Rice)
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Author:
Date:
2016-12-20
[Abstract] Superoxide ions (O2-) and hydrogen peroxide (H2O2) are the reactive oxygen species (ROS) that play a significant role in regulation of many plant processes. The level of O2- ions is determined qualitatively using nitrobluetetrazolium (NBT) assay while the H2O2 is qualitatively estimated using 3,3-diaminobenzidine (DAB) and 2’,7’-dichlorodihydrofluorescein diacetate (H2DCFDA) assay. Further the aqueous content of H2O2 is estimated quantitatively using ferrous oxidation-xylenol orange (FOX) assay.
[摘要] 超氧化物离子（O 2 - ）和过氧化氢（H 2 O 2 O 2）是起重要作用的活性氧（ROS）在许多植物过程的调节。使用硝基四氮唑（NBT）测定定性地测定O 2 - 离子的水平，同时使用3,3 - 二氨基联苯胺（DAB）和2'，7'-二氯二氢荧光素二乙酸酯（H 2 N 2 DCFDA）测定。此外，使用亚铁氧化 - 二甲苯酚橙（FOX）测定定量地估计H 2 O 2 O 2的含水量。
背景 超氧化物离子和过氧化氢是在许多情况下起主要作用的活性氧分子涉及植物生长发育的过程包括非生物胁迫耐受性。为了更好地了解ROS对这些过程的调节，对不同类型ROS的定性和定量估计具有重要意义。通过由NADPH氧化酶系统介导的电子从NADPH转移到氧（O 2），产生O 2。使用NBT测定法在水稻幼苗中估计这些离子，其基于通过O将黄色NBT还原成深蓝色不溶性甲the的原理（Kaur等人，2016）。
＆nbsp; H 2是另一种作为调节不同植物过程的重要信号分子的活性氧分子。在使用DAB和H 2 DCFDA测定的水稻幼苗中定性地估计H 2 O 2 O 2的含量（Kaur等人，2016）。 DAB测定是基于DAB与H 2 O 2 O 2的反应形成深棕色聚合产物的原理，而H 2 N 2 ...
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| Isolation of Heterocysts from Anabaena sp. PCC 7120
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Author:
Date:
2015-04-20
[Abstract] During combined nitrogen step-down, filaments of cyanobacterium Anabaena sp. PCC 7120 differentiate about 5-10% of vegetative photosynthetic cells into heterocysts, the specialized cells for N2 fixation (Walk, 1996). Heterocysts have a thick cell wall reducing permeation of O2 and consist of two additional layers composed of glycolipids and polysaccharides. The difference in structure and composition of the cell wall between heterocysts and vegetative cells allows separation and isolation of heterocyst. Heterocysts isolated by this protocol can be subjected to protein analysis and activity measurements, which do not require strict anaerobic conditions.
[摘要] 在组合氮降压期间，蓝细菌的鱼腥藻属 PCC 7120将约5-10％的营养光合细胞分化成异种细胞，这是用于N sub2固定的特化细胞（Walk，1996）。 杂合体具有厚的细胞壁，减少O 2的渗透，并且由两个由糖脂和多糖组成的附加层组成。 杂合细胞和营养细胞之间的细胞壁的结构和组成的差异允许异种细胞的分离和分离。 通过该方案分离的杂合体可以进行蛋白质分析和活性测量，其不需要严格的厌氧条件。
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| Purification and Detection of a PDGA Depolymerase from Pusillimonas noertemannii
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Author:
Date:
2014-11-05
[Abstract] The purification of a target protein from a complex mixture of proteins is a challenging undertaking. If the target protein has been previously characterised, then information such as subcellular location, function, molecular weight and pI can be used for the design of a purification strategy. However, if the target protein is uncharacterised or little information regarding its characteristics is available, a generic purification protocol can be employed that is optimised as additional characteristics of the target protein are determined during subsequent purification steps. Herein, we describe the protocol for the purification and detection of a poly-γ-D-glutamic acid (PDGA) depolymerase from a consortium culture of two Gram-negative bacteria using a combination of chromatography, ...
[摘要] 从蛋白质的复杂混合物中纯化靶蛋白是一项具有挑战性的任务。 如果靶蛋白以前已被表征，则诸如亚细胞定位，功能，分子量和pI的信息可用于设计纯化策略。 然而，如果目标蛋白质是未表征的或者关于其特征的很少的信息可用，则可以使用通用的纯化方案，其随着在随后的纯化步骤期间测定靶标蛋白质的附加特征而被优化。 在这里，我们描述了从两个革兰氏阴性细菌的联合培养物使用色谱，2D电泳和酶谱的组合纯化和检测聚-γ-D-谷氨酸（PDGA）解聚酶的协议。
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