| Detection of Nitric Oxide and Determination of Nitrite Concentrations in Arabidopsis thaliana and Azospirilum brasilense
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Author:
Date:
2016-03-20
[Abstract] There is now general agreement that nitric oxide (NO) is an important and almost ubiquitous signal in plants. Nevertheless, there are still many controversial observations and differing opinions on the importance and functions of NO in plants. Partly, this may be due to the difficulties in detecting and quantifying NO. Here, we summarize protocols for detecting NO and quantifying nitrite concentration in Arabidopsis seedlings. We also present a method to measure NO in biofilms formed by the plant growth promoting rhizobacteria Azospirillum brasilense (A. brasilense). NO in oxygen-containing aqueous solutions has a short half-life that is often attributed to a rapid oxidation to nitrite. Here we detail the use of the fluorescent probe DAF-FM DA and the ...
[摘要] 现在普遍认为一氧化氮（NO）是植物中重要的和几乎普遍存在的信号。然而，对于植物中NO的重要性和功能仍有许多有争议的观察和不同的观点。部分地，这可能是由于检测中的困难以及甚至更多的在定量NO。在这里，我们总结了在拟南芥幼苗中检测NO和定量亚硝酸盐浓度的方案以及在由植物生长促进根瘤菌（Azospirillum brasilense）形成的生物膜中的NO实时测量（ > A。brasilense ）。含氧水溶液中的NO具有短的半衰期，其通常归因于快速氧化成亚硝酸盐。在这里我们详细的使用荧光探针DAF-FM DA和电化学方法分别直接检测和定量NO，Griess试剂通过其氧化的亚硝酸盐形式间接检测NO。这些方案可用于多种细胞类型和植物的不同组织以及微生物。
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| Construction of Chromosomal In-Frame Deletion Mutants in Rhodobacter capsulatus
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Author:
Date:
2013-02-20
[Abstract] Markerless chromosomal in-frame deletion mutants are often used to avoid polar effects, conserve antibiotic resistance markers or make multiple mutations. This protocol capitalizes on the sacB gene (lethal in the presence of sucrose), for selection against a single cross-over event, on a suicide vector (contains no Rhodobacter capsulatus compatible origin of replication). The bacterial strains, media and suicide vectors in this protocol are specific for R. capsulatus, but the concepts can be applied to other species.
[摘要] 无记号染色体框内缺失突变体常用于避免极性效应，保存抗生素抗性标记或产生多个突变。 该协议利用了在sacC基因（在蔗糖存在下致死），用于针对单个交叉事件的选择，在自杀载体上（不含有红细菌Rhodobacter capsulatus 兼容性 复制起点）。 该方案中的细菌菌株，培养基和自杀载体对于R是特异性的。 capsulatus ，但这些概念可应用于其他物种
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| Rhodobacter capsulatus Gene Transfer Agent Transduction Assay
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Author:
Date:
2013-02-20
[Abstract] The gene transfer agent (GTA) is a bacteriophage-like particle that transfers genomic DNA from a donor to a recipient bacterium. The Rhodobacter capsulatus GTA (RcGTA) was the first to be studied and this protocol has been optimized for RcGTA transduction, although it could be modified for other bacteria containing a GTA. The RcGTA transduction assay can be used to determine transduction efficiencies, to create gene knock-outs, or to create new strains by transferring alleles from one strain to another.
[摘要] 基因转移剂（GTA）是将基因组DNA从供体转移到受体细菌的噬菌体样颗粒。 红细菌（Rhodobacter capsulatus）GTA（RcGTA）是第一个被研究的，并且该方案已经针对RcGTA转导进行了优化，尽管其可以对于包含GTA的其他细菌进行修饰。 RcGTA转导测定可用于测定转导效率，产生基因敲除，或通过将等位基因从一个菌株转移到另一个菌株来产生新菌株。
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