| RNA Isolation from Synechocystis
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Author:
Date:
2015-03-20
[Abstract] The protocol describes the procedure of total RNA isolation from cells of the cyanobacterium Synechocystis sp. PCC 6803. This protocol is also applicable to Synechococcus elongatus PCC 7942 and PCC 6301, Thermosynechococcus vulcanus, and other unicellular and filamentous species of cyanobacteria that do not have thick polysaccharide-containing outer layers. For the latter, Trizol-containing protocols should be adapted. The yield of RNA depends on optical density of cyanobacterial culture and may reach up to 10-20 µg of total RNA per 1 ml of cell culture. RNA isolated by this method can be used for Northern blot hybridization, RT-qPCR, microarrays and Next Generation Sequencing.
[摘要] 该方案描述了从蓝细菌集胞藻的细胞总RNA分离的程序。 PCC 6803。该方案也适用于细长聚球藻PCC 7942和PCC 6301，硫酸高热杆菌（Thermosynechococcus vulcanus）和其它单细胞和丝状物种的蓝细菌，其不具有含多糖的 外层。 对于后者，应该修改含有Trizol的协议。 RNA的产量取决于蓝细菌培养物的光密度，并且可以达到每1ml细胞培养物高达10-20μg的总RNA。 通过该方法分离的RNA可用于Northern印迹杂交，RT-qPCR，微阵列和下一代测序。
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| Macromolecular Biosynthesis Assay for Evaluation of Influence of an Antimicrobial on the Synthesis of Macromolecules
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Author:
Date:
2013-06-20
[Abstract] One of the most compelling approaches in the discovery of novel antimicrobials is screening of natural sources. In our publication we report on the activity of a compound 8-hydroxyserrulat-14-en-19-oic acid (EN4), a diterpene isolated from the Australian plant Eremophila neglecta. We evaluate its applicability for treatment of implant-associated infections. A comprehensive analysis of the mechanism of action of EN4 against staphylococci revealed its membranolytic properties and a general inhibition of macromolecular biosynthesis, which was confirmed in a macromolecular biosynthesis assay and suggested a multitarget activity. The method used to investigate an influence of EN4 on the synthesis of peptidoglycan, RNA, DNA and proteins is based on precipitation of macromolecules with ...
[摘要] 在发现新的抗微生物剂中最引人注目的方法之一是筛选天然来源。在我们的出版物中，我们报告了一种化合物8-羟基胆固醇-14-烯-19-酸（EN4）（一种从澳大利亚植物中分离的二萜烯）的活性。我们评估其适用于植入物相关感染的治疗。对EN4对葡萄球菌的作用机制的综合分析揭示其膜分解性质和大分子生物合成的一般抑制，其在大分子生物合成测定中被证实并提出多靶点活性。用于研究EN4对肽聚糖，RNA，DNA和蛋白质的合成的影响的方法是基于大分子与三氯乙酸的沉淀。这些大分子由相应的[3 H] - 标记的前体合成。测量掺入和不掺入抗微生物剂的放射性，并反映测试化合物的作用模式。使用具有已知作用机制的抗生素作为对照。
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