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Fisher BioReagentsTM Powdered Agar

Fisher生物试剂TM粉状琼脂

Company: Thermo Fisher Scientific
Catalog#: BP1423-2
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Transfer of Large Contiguous DNA Fragments onto a Low Copy Plasmid or into the Bacterial Chromosome
Author:
Date:
2016-11-20
[Abstract]  Bacterial pathogenicity islands and other contiguous operons can be difficult to clone using conventional methods due to their large size. Here we describe a robust 3-step method to transfer large defined fragments of DNA from virulence plasmids or cosmids onto smaller autonomously replicating plasmids or directly into defined sites in the bacterial chromosome that incorporates endogenous yeast and λ Red homologous recombination systems. This methodology has been successfully used to isolate and integrate at least 31 kb of contiguous DNA and can be readily adapted for the recombineering of E. coli and its close relatives. [摘要]  由于其大尺寸,细菌致病性岛和其它连续操纵子可能难以使用常规方法克隆。在这里我们描述了一个强大的3步法从毒力质粒或粘粒转移大型定义的片段到更小的自主复制质粒或直接到细胞染色体,并入内源性酵母和λ红色同源重组系统的定义网站。该方法已经成功地用于分离和整合至少31kb的连续DNA,并且可以容易地适应于E的重组。大肠杆菌及其近亲。

[背景] 分离和繁殖大片DNA的能力大大扩展了基因网络和操纵子的研究。然而,用于该目的的传统使用的工程质粒,例如细菌人工染色体(BAC),虽然极其有用,但是受到DNA稳定性,拷贝数和复杂装配要求的问题的限制。或者,将构建体直接并入细菌染色体中通过减少由于存在多个基因拷贝引起的基因表达的变化以及确保基因的稳定维持而提供了优点,同时还避免了对抗生素选择的需要。这里描述的方法最初被设计为捕获和转移编码Shigella flexneri 3型分泌系统的31kb DNA操纵子到大肠杆菌染色体上(Reeves ...

Water Deficit Treatment and Measurement in Apple Trees
Author:
Date:
2015-02-05
[Abstract]  Water is considered perhaps the most limiting factor for plant growth and productivity (Boyer, 1982), and climate change predicts more frequent, more severe and longer drought periods for a significant portion of the world in coming years. Unfortunately, drought resistance is particularly difficult to measure due in part to the complexity of the underlying biology that contributes to a plant’s ability to cope with water limitations. For example, water deficit is frequently examined by detaching leaves or withholding water for a set period of time prior to tissue collection. Such approaches may elucidate the early stages of drought response but are generally not physiologically relevant for maintenance of drought resistance over a longer period. A more realistic approach is to impose a ... [摘要]  水被认为可能是植物生长和生产力的最大限制因素(Boyer,1982),气候变化预测未来几年世界上相当大一部分地区的更频繁,更严重和更长的干旱期。不幸的是,抗旱性特别难以测量,部分原因是基础生物学的复杂性,有助于植物应对水分限制的能力。例如,在组织收集之前,通常通过将叶子或保留水分离一段设定的时间来检查水缺乏。这些方法可以阐明干旱反应的早期阶段,但是通常在较长时期内维持抗旱性不是生理学相关的。更现实的方法是用持续的土壤水分水平强加逐步的水限制,特别是在多年生木本的情况下。我们在这里描述一个协议,在受控的实验室条件下施加长期缺水,允许分子生物学方法来了解木本植物如何生存的严重水限制。代表性数据可以在Artlip等人(1997)和Bassett等人(2014)中找到。

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