| Determination of the Predatory Capability of Bdellovibrio bacteriovorus HD100
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Author:
Date:
2017-03-20
[Abstract] Bdellovibrio bacteriovorus HD100 is an obligate predator that preys upon a wide variety of Gram negative bacteria. The biphasic growth cycle of Bdellovibrio includes a free-swimming attack phase and an intraperiplasmic growth phase, where the predator replicates its DNA and grows using the prey as a source of nutrients, finally dividing into individual cells (Sockett, 2009). Due to its obligatory predatory lifestyle, manipulation of Bdellovibrio requires two-member culturing techniques using selected prey microorganisms (Lambert et al., 2003). In this protocol, we describe a detailed workflow to grow and quantify B. bacteriovorus HD100 and its predatory ability, to easily carry out these laborious and time-consuming techniques.
[摘要] HDD是一种专有捕食者，可以用于各种革兰氏阴性菌。 Bdellovibrio的双相生长周期包括自由游泳攻击阶段和胞内生长期，其中捕食者复制其DNA并使用猎物作为营养源，最终分成单个细胞（Sockett ，2009）。由于其必要的掠夺性生活方式，使用选择的猎物微生物（Lambert等人，2003）的双重培养技术的操作需要Bdellovibrio 。在这个协议中，我们描述了一个详细的工作流程来增长和量化。细菌性HD100及其掠夺能力，轻松实施这些费时费力的技术。
在过去几年中，Bdellovibrio 已经吸引了科学界的兴趣，并且已经开发了若干应用，如进化研究（Davidov和Jurkevitch，2009），鉴定新的生物催化剂（Martínezet al。 （2012）），治疗应用（Atterbury等人，2011），或使用Bdellovibrio的生物技术应用作为回收附加值的溶解剂细胞内生物产物（Martínez等人，2016）。由于对Bdellovibrio 的兴趣日益增加，已经开发了用于量化这种掠食性细菌的不同间接方法（Mahmoud等人，2007; Lambert和Sockett，2008; Van Essche et al。，2009）。然而，通过双层方法直接量化B。o is is is is is is is is is to to to to ...
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| Quantification of Chitinase Activity in Fusarium oxysporum
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Author:
Date:
2016-08-05
[Abstract] Fungal morphogenetic development requires modification and plasticity of the cell wall, which implies synthesis and remodelling of its components, including chitin and glucan. Thus chitinase and glucanase activities are crucial for cell-wall biogenesis and cell division. Quantification of chitinase activity might be useful to identify structural defects that could negatively influence growth and morphogenesis of some filamentous fungi like Fusarium oxysporum, which produces both intracellular and secreted chitinases. The chitinolytic enzymes are categorized based on their enzymatic action on chitin substrates. Endochitinases are defined as the enzymes catalyzing the random cleavage at internal points in the chitin chain. Exochitinases catalyze the progressive release of ...
[摘要] 真菌形态发生需要细胞壁的修饰和可塑性，这意味着其组分（包括壳多糖和葡聚糖）的合成和重塑。因此几丁质酶和葡聚糖酶活性对于细胞壁生物发生和细胞分裂是至关重要的。几丁质酶活性的定量可能有助于鉴定可能负面影响一些丝状真菌如产生细胞内和分泌型几丁质酶的尖孢镰刀菌的生长和形态发生的结构缺陷。基于它们对壳多糖底物的酶作用将壳聚糖分解酶分类。内酯酶被定义为催化几丁质链中内部点的随机裂解的酶。外切球蛋白酶催化乙酰壳寡糖或N-乙酰葡糖胺从几丁质的非还原端逐步释放，因此分别称为壳聚糖酶和β-N-乙酰氨基葡糖苷酶。在这里，我们描述了一种简单的方法来轻易地纯化几丁质酶，以便比较不同的F的内切几丁质酶活性和外切几丁质酶活性。尖孢镰孢菌株。该方案可适用于任何真菌物种。
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| Determination of Intra- and Extracellular Glucose in Mycelium of Fusarium oxysporum
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Author:
Date:
2016-07-20
[Abstract] To study alterations in the metabolism and/or in the transport of glucose during Fusarium oxysporum vegetative growth, we determined intracellular glucose levels in different fungal strains, as well as the amount of glucose remaining in the supernatants after growth in synthetic medium (SM) supplemented with either 0.05 or 2.5% glucose. We used the Glucose (GO) Assay Kit (Sigma-Aldrich) following the instructions of the manufacturer with some modifications. The protocol described here can be applied to other filamentous fungi.
[摘要] 为了研究尖镰孢营养生长期间葡萄糖的代谢和/或转运中的改变，我们测定了不同真菌菌株中的细胞内葡萄糖水平以及生长后上清液中剩余的葡萄糖量 在补充有0.05或2.5％葡萄糖的合成培养基（SM）中。 我们使用葡萄糖（GO）测定试剂盒（Sigma-Aldrich）按照制造商的指示进行一些修改。 这里描述的协议可以应用于其他丝状真菌。
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