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Glass bottles 5 L

Company: Kavalierglass of North America
Catalog#: CFB020
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Isolation of Ribosomal Particles from the Unicellular Cyanobacterium Synechocystis sp. PCC 6803
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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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