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Company: BD
Catalog#: 307736
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Bacterial Competition Assay Based on Extracellular D-amino Acid Production
Author:
Date:
2018-04-05
[Abstract]  Bacteria live in polymicrobial communities under tough competition. To persist in a specific niche many species produce toxic extracellular effectors as a strategy to interfere with the growth of nearby microbes. One of such effectors are the non-canonical D-amino acids. Here we describe a method to test the effect of D-amino acid production in fitness/survival of bacterial subpopulations within a community. Co-cultivation methods usually involve the growth of the competing bacteria in the same container. Therefore, within such mixed cultures the effect on growth caused by extracellular metabolites cannot be distinguished from direct physical interactions between species (e.g., T6SS effectors). However, this problem can be easily solved by using a filtration unit that allows free ... [摘要]  在激烈的竞争中,细菌生活在多种微生物群落中。为了坚持特定的生态位,许多物种会产生有毒的细胞外效应物作为干扰附近微生物生长的策略。这种效应子之一是非规范的D-氨基酸。在这里我们描述一种方法来测试D-氨基酸生产对社区内细菌亚群的适应/存活的影响。共培养方法通常涉及相同容器中竞争细菌的生长。因此,在这种混合培养物中,细胞外代谢物对生长的影响不能与物种间的直接物理相互作用区分开(例如T6SS效应物)。然而,通过使用允许小分解代谢物(例如L-和D-氨基酸)自由扩散的过滤单元可以容易地解决这个问题,同时将不同亚群保持在独立区室中。

通过这种方法,我们已经证明D-精氨酸是由霍乱弧菌产生的杀菌剂效应物,其强烈影响不同微生物亚群的存活。此外,D-精氨酸可作为混合菌群中的一种协同工具,用于保护非生产成员免受竞争细菌的侵害。

【背景】细菌生活在多种多样的物种共存并争夺现有资源的多种微生物群落中。细菌设计为在特定生态位持续存在的许多策略之一是产生有毒的细胞外代谢物作为干扰其他微生物生长和/或生存力的策略。已知D-氨基酸长时间在细菌培养物中具有细胞形状和活力的强大作用(Bopp,1965; Fox等人,1944; Kobayashi等人, 1948年; Yaw和Kakavas,1952年; Lark和Lark,1959年; Grula,1960年; ...

Determination of the Predatory Capability of Bdellovibrio bacteriovorus HD100
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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