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Company: Duchefa Biochemie
Catalog#: M0231
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Multiplexed GuideRNA-expression to Efficiently Mutagenize Multiple Loci in Arabidopsis by CRISPR-Cas9
[Abstract]  Since the discovery of the CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein (Cas) as an efficient tool for genome editing in plants (Li et al., 2013; Shan et al., 2013; Nekrasov et al., 2013), a large variety of applications, such as gene knock-out, knock-in or transcriptional regulation, has been published. So far, the generation of multiple mutants in plants involved tedious crossing or mutagenesis followed by time-consuming screening of huge populations and the use of the Cas9-system appeared a promising method to overcome these issues. We designed a binary vector that combines both the coding sequence of the codon optimized Streptococcus pyogenes Cas9 nuclease under the control of the Arabidopsis thaliana ... [摘要]  自从发现CRISPR(聚集的定期交织的短回文重复) - 相关蛋白(Cas)作为植物基因组编辑的有效工具(Li等人,2013; Shan等人已经出版了诸如基因敲除,敲入或转录调控等各种各样的应用,例如,2013; Nekrasov等人,2013)。到目前为止,植物中多种突变体的产生涉及繁琐的杂交或诱变,随后大量人群的耗时筛选,Cas9系统的使用似乎是有希望的方法来克服这些问题。我们设计了一种二元载体,其结合了在拟南芥UBIQUITIN10(UBQ10)启动子和引导RNA(gRNA)控制下的优化的化脓性链球菌(Caspase)密码子的编码序列)由 A驱动的表现盒。拟南芥U6 - 启动子,用于在拟南芥中进行有效的多重编辑(阎等人,2016年)。在这里,我们描述了一个逐步的方案,以经济有效的方式生成含有多个gRNA的二元载体和基于经典克隆方法的Cas9核酸酶。背景 RNA引导的Cas9系统源于针对外源DNA的细菌防御系统(Sorek等人,2013)。由于其高效率,易于处理和多重编辑的可能性,已经被认为是基因组编辑的选择方法。通常,Cas9基因编辑系统涉及单个合成RNA分子,其指导Cas9蛋白质靶向所需DNA位点以进行基因组修饰或转录控制的gRNA。 gRNA-Cas9复合物通过gRNA-DNA配对识别靶向的DNA,并需要存在原始相邻基序(PAM)。 ...

An Assay to Study Botrytis cinerea-infected Grapevine Leaves Primed with Pseudomonas fluorescens
[Abstract]  Grapevine (Vitis vinifera L.) is susceptible to an array of diseases among them the grey mold caused by the necrotrophic fungus Botrytis cinerea that decreases grape productivity and quality. To ensure a satisfactory yield and harvest quality numerous chemical fungicides are required, but they have serious drawbacks. One alternative is the use of beneficial bacteria to improve plant health. Pseudomonas fluorescens has been shown to trigger a plant-mediated resistance response in aboveground plant tissues against fungal, oomycete, bacterial, and viral pathogens. Triggered plant resistance exploits mechanisms of the plant immune system through a priming state that provides plants with enhanced capacity for rapid and strong activation of defense responses after ... [摘要]  葡萄(葡萄(Vitis vinifera)L.)易受一系列疾病的影响,其中包括由坏死性真菌葡萄孢菌引起的灰霉病,其降低葡萄生产力和质量。为了确保令人满意的产量和收获质量,需要许多化学杀真菌剂,但它们具有严重的缺点。一个替代方案是使用有益细菌来改善植物健康。已经显示荧光假单胞菌在地上植物组织中触发植物介导的对真菌,卵菌,细菌和病毒病原体的抗性反应。触发的植物抗性通过启动状态利用植物免疫系统的机制,其提供植物在病原体感染后快速和强烈地激活防御反应的增强能力,导致较低的适合度成本。有益细菌的引发应答包括防御相关基因的诱导表达,细胞壁增强和病原体感染后次生代谢产物的产生。在该方案中,我们描述了根据Verhagen评价有益细菌荧光假单胞菌PTA-CT2对葡萄植​​物的引发状态及其对灰葡萄孢的抗性水平的实验设计等。 (2011)和Gruau等人。 (2015)。

Salinity Assay in Arabidopsis
[Abstract]  Salinity is an important environmental constraint to crop productivity in arid and semi-arid regions of the world. The evaluation of the responses to salinity of different Arabidopsis ecotypes or transgenic lines is important to identify and investigate the role of different key genes. These new characterized genes involved in the response to salinity stress are of great interest to be incorporated in crops breeding programs. Here we provide a reproducible method to evaluate the performance of Arabidopsis lines to salinity stress by analysing primary and lateral root growth and fresh weight of plants grown under in vitro conditions in growth chambers. Even though NaCl is the most frequent used salinity tests, other salts (e.g. KCl, MgCl2) can ... [摘要]  盐度是世界干旱和半干旱地区作物生产力的重要环境约束。对不同拟南芥生态型或转基因株系的盐度的响应的评价对于鉴定和研究不同关键基因的作用是重要的。这些新的表征基因参与对盐分胁迫的反应是非常感兴趣的被纳入作物育种计划。在这里我们提供了一种可再现的方法,通过分析在生长室中的体外条件下生长的植物的初级和侧根生长和鲜重来评估拟南芥株系对盐度胁迫的性能。 。即使NaCl是最常用的盐度测试,也可以通过该方法评价其它盐(例如KCl,MgCl 2)。拟南芥植物在这些条件下可以维持15-20天,但是在前10天内可以观察到对生长和生物量的影响,这取决于所使用的盐和浓度。