| Conjugation Protocol Optimised for Roseburia inulinivorans and Eubacterium rectale
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Author:
Date:
2020-04-05
[Abstract] Roseburia and Eubacterium species of the human gut microbiota play an important role in the maintaince of human health, partly by producing butyrate, the main energy source of our colonic epithelial cells. However, our knowledge of the biochemistry and physiology of these bacteria has been limited by a lack of genetic manipulation techniques. Conjugative transposons previously introduced into Roseburia species could not be easily modified, greatly limiting their applicability as genetic modification platforms. Modular plasmid shuttle vectors have previously been developed for Clostridium species, which share a taxonomic order with Roseburia and Eubacterium, raising the possibility that these vectors could be used in these organisms. ...
[摘要] [摘要 ] 人体肠道菌群中的玫瑰菌属和真细菌属在维持人类健康中起着重要作用，部分原因是产生丁酸盐，这是我们结肠上皮细胞的主要能源。但是，由于缺乏基因操作技术，我们对这些细菌的生物化学和生理学的认识受到限制。先前引入玫瑰花属物种的共轭转座子不容易被修饰，极大地限制了它们作为基因修饰平台的适用性。MOD ular质粒穿梭载体先前已经开发了用于梭菌物种，其与共享一个分类次序ř oseburia 和真杆菌，提高这些矢量可以在这些生物体中使用的可能性。在这里，我们描述了一种优化缀合协议使得能够自主复制的质粒的从转印大肠杆菌供体菌株为罗斯氏inulinivorans 和真杆菌rectale 。质粒的模块性质及其通过自主复制在受体细菌中得以维持的能力使其成为研究异源基因表达的理想之选，并成为其他遗传工具（包括反义RNA沉默或II 型移动子中断子基因破坏策略）的平台。
[背景 ] 玫瑰菌和真细菌属人类肠道菌群中含量最高的细菌（Zhernakova 等，2016），它们通过利用饮食和宿主衍生的多糖影响人类健康（Scott 等，2006和2011； Cockburn 等） 。，2015 ; 谢里登等人，2016 ）并产生促进健康的代谢物丁酸作为发酵终产物（邓肯等人，2002和2006） 。另外，这些物种能够通过鞭毛调节宿主免疫（Neville ...
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| In vitro Assay of the Glycosyltransferase Activity of a Heterologously Expressed Plant Protein
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Author:
Date:
2014-11-05
[Abstract] Glycosyltransferases are carbohydrate active enzymes containing catalytic modules involved in catalysing the biosynthesis of glycosidic bonds in oligo- and polysaccharides and glycoconjugates. One of the most comprehensive collections of Carbohydrate Active enZYmes is the CAZy database (http://www.cazy.org) comprising 120,000 glycosyltransferases allocated to 96 families based mainly on sequence homologies of their conserved and catalytically active domains (Cantarel et al., 2009). Interestingly, the glycosyltransferase activities of only about 1.6% of these proteins have been experimentally characterized (Lombard et al., 2014). In recent years, membrane-bound glycosyltransferases of a number of families have been shown ...
[摘要] 糖基转移酶是含有催化模块的碳水化合物活性酶，其涉及催化寡糖和多糖和糖缀合物中糖苷键的生物合成。 Carbohydrate Active enZYmes最全面的集合之一是CAZy数据库（ http://www.cazy.org ） ），其包含分配给96个家族的120,000个糖基转移酶，主要基于其保守和催化活性结构域的序列同源性（Cantarel等人，2009）。有趣的是，仅约1.6％的这些蛋白质的糖基转移酶活性已经通过实验表征（Lombard等人，2014）。近年来，已显示许多家族的膜结合糖基转移酶在植物细胞壁多糖的生物合成中起关键作用（Doblin等人，2010; Scheller和Ulvskov，2010 ; ...
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