| Isolation of Ribosomal Particles from the Unicellular Cyanobacterium Synechocystis sp. PCC 6803
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Author:
Date:
2017-03-20
[Abstract] Isolation of ribosomal particles is an essential step in the study of ribosomal components as well as in the analysis of trans-acting factors that interact with the ribosome to regulate protein synthesis and modulate the expression profile of the cell in response to different environmental conditions. In this protocol, we describe a procedure for the isolation of 70S ribosomes from the unicellular cyanobacterium Synechocystis sp. PCC 6803 (hereafter Synechocystis). We have successfully used this protocol in our study of the cyanobacterial ribosomal-associated protein LrtA, which is a homologue of bacterial HPF (hibernation promoting factor) (Galmozzi et al., 2016).
[摘要] 核糖体颗粒的分离是研究核糖体组分以及与核糖体相互作用以调节蛋白质合成并调节细胞表达谱的反式因子的分析中必不可少的步骤响应不同的环境条件。在本协议中，我们描述了从单细胞蓝细菌集胞藻分离70S核糖体的过程。 PCC 6803（以下简称集胞藻）。我们已经成功地使用这个方案来研究蓝细菌核糖体相关蛋白LrtA，它是细菌HPF（冬眠促进因子）的同系物（Galmozzi等人，2016）。
背景 据报道蓝藻核糖体几乎没有生物化学研究。已经通过差速离心分离70S核糖体颗粒，然后通过二维电泳分析核糖体蛋白质（Sato et al。，et al。 ，1998）。核糖体也已经从聚球藻（Spechococcus）进行制备。 PCC 6301细胞使用组合差速离心和蔗糖步骤梯度的方案（Sugita等人，2000）。通过差速离心分离细胞提取物也被用于制备核糖体样品用于在不同的聚球藻菌株中开发体外翻译系统（Mutsuda和Sugiura，2006 ）。基于针对聚球藻（Sugita等人，2000）所述的方法，本文所述的针对集胞藻的方法允许使用以下方式纯化核糖体颗粒线性蔗糖梯度的超速离心。
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| Testing the Effect of UV Radiation on the Survival of Burkholderia glumae
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Author:
Date:
2016-03-05
[Abstract] Burkholderia glumae (B. glumae) is becoming a serious threat in the major rice producing areas of the world. It was reported that Burkholderia spp., including B. glumae, are adapted to a wide range of ecological niches. Different bacterial strains show different levels of UV tolerance which may be due to the presence of different protection mechanisms. Previously we reported that pigment producing strains of B. glumae are more tolerant to UV radiation than non-pigmented strains. Here, we describe the protocol of UV tolerance assay for B. glumae in different exposure times. Using this protocol, we can calculate the survival rate of B. glumae strains, as well as other bacterial species, in exposure to UV radiation.
[摘要] Burkholderia glumae （ B。glumae ）正在成为世界主要稻米生产地区的严重威胁。 据报道，伯克霍尔德菌属包括 B。 腮腺炎，适应广泛的生态位。 不同的细菌菌株显示不同水平的UV耐受性，这可能是由于存在不同的保护机制。 以前，我们报告了颜色产生菌株的B。 腮腺炎比非色素菌株更耐紫外线辐射。 在这里，我们描述了紫外线耐受性测定的方案。 腮腺炎在不同的暴露时间。 使用这个协议，我们可以计算的生存率。 腮腺炎菌株，以及其他细菌物种，暴露于紫外线辐射。
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| Airbrush Infiltration Method for Pseudomonas syringae Infection Assays in Soybean
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Author:
Date:
2015-03-20
[Abstract] We developed this protocol to assay the extent of proliferation of Pseudomonas syringae pv. glycinea in soybean leaves. This method specifically enables accurate pathogenesis assays of soybean plants at V2/V3 (2nd/3rd trifoliate) or higher stages of growth. The leaves of soybean plants at these growth stages are not amenable to bacterial infiltration using routine needleless syringe infiltration due to the high number of trichomes on these mature leaves. This method enables efficient infiltration of bacteria into the epidermal cells of mature leaves using a pressure pump.
[摘要] 我们开发了该方案来测定丁香假单胞菌pv的增殖程度。 在大豆叶中。 该方法特异性地能够在V2/V3（2期/3期/3期/3期/3期/3期）或更高生长阶段进行大豆植物的准确发病机理测定。 由于这些成熟叶上的大量毛状体，大豆植物在这些生长阶段的叶子不适于使用常规无针注射器浸润的细菌浸润。 该方法使用压力泵能够有效地将细菌浸润到成熟叶的表皮细胞中。
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