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Falcon® 50 mL High Clarity PP Centrifuge Tube, Conical Bottom, Sterile

Falcon ® 50mL高清PP离心管,锥形底

Company: Corning
Catalog#: 352070
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Trimolecular Fluorescence Complementation (TriFC) Assay for Direct Visualization of RNA-Protein Interaction in planta
Author:
Date:
2017-10-20
[Abstract]  RNA-Protein interactions play important roles in various eukaryotic biological processes. Molecular imaging of subcellular localization of RNA/protein complexes in plants is critical for understanding these interactions. However, methods to image RNA-Protein interactions in living plants have not yet been developed until now. Recently, we have developed a trimolecular fluorescence complementation (TriFC) system for in vivo visualization of RNA-Protein interaction by transient expression in tobacco leaves. In this method, we combined conventional bimolecular fluorescence complementation (BiFC) system with MS2 system (phage MS2 coat protein [MCP] and its binding RNA sequence [MS2 sequence]) (Schonberger et al., 2012). Target RNA is tagged with 6xMS2 and MCP and RNA binding ... [摘要]  RNA-蛋白质相互作用在各种真核生物过程中起重要作用。 RNA /蛋白质复合物在植物中亚细胞定位的分子成像对于理解这些相互作用至关重要。然而,到目前为止,尚未开发在活植物中形成RNA-蛋白质相互作用的方法。最近,我们开发了一种三分子荧光互补(TriFC)系统,用于在烟草叶中瞬时表达的RNA-蛋白质相互作用的体内可视化。在这种方法中,我们将传统的双分子荧光互补(BiFC)系统与MS2系统(噬菌体MS2外壳蛋白[MCP]及其结合RNA序列[MS2序列])(Schonberger等人,2012)相结合, 。目标RNA用6xMS2标记,MCP和RNA结合蛋白与YFP片段融合。编码这种融合RNA和蛋白质的DNA构建体用土壤杆菌悬浮液渗入烟草叶中。通过共焦显微镜观察体内的RNA-蛋白质相互作用
【背景】近来,多种类型的长非编码RNA(lncRNA)已经被鉴定并显示出在转录调节和染色质修饰中起重要作用(St Laurent等人,2015)。到目前为止,lncRNA介导的功能的大多数分子机制与RNA-蛋白质相互作用密切相关(St ...

In vitro Co-culture of Mesenchymal Stem Cells and Endothelial Colony Forming Cells
Author:
Date:
2017-10-20
[Abstract]  The discovery of endothelial colony forming cells (ECFCs) with robust self-renewal and de novo vessel formation potentials suggests that ECFCs can be an excellent cell source for cardiovascular diseases treatment through improving neovascularization in the ischemic tissues. However, their engraftment after transplantation resulted to be low. Previous studies showed mesenchymal stem/stromal cells (MSCs) could improve the survival and capillary formation capacity of ECFCs in co-culture systems. In this article, we describe a protocol for in vitro co-culture of MSCs and ECFCs to prime ECFCs for better engraftment. [摘要]  发现具有强大自我更新和从头血管形成潜力的内皮细胞集落形成细胞(ECFCs)表明,ECFC可以通过改善缺血组织的新生血管形成,成为心血管疾病治疗的优良细胞来源。 然而,移植后的移植导致了低位移植。 以前的研究显示间充质干/基质细胞(MSC)可以改善共培养系统中ECFCs的存活和毛细管形成能力。 在这篇文章中,我们描述了体外协调MSCs和ECFCs共同培养ECFC以实现更好的移植。
【背景】内皮祖细胞(EPC)被定义为能够通过血管发生过程形成新血管的细胞群。 2004年,Ingram等人鉴定了来自人脐带血的称为“内皮细胞集落形成细胞(ECFC)”的离体培养物中的特异性高度增殖的EPC群体Ingram等人,2004),并且这些细胞最近被宣布代表EPCs(Medina等人,2017)。类似的群体也可以从具有等效血管化潜力和临床相关数量的人类胎盘组织中分离(Patel等人,2013; Shafiee等人,2015) )。因此,ECFC移植已被提出作为缺血性疾病如心肌梗塞或关键性腿部缺血的治疗方法。然而,移植后的ECFCs植入物和血管生成潜力被证明是低的(Shafiee等人,2017; ...

Protein Expression and Purification of the Hsp90-Cdc37-Cdk4 Kinase Complex from Saccharomyces cerevisiae
Author:
Date:
2017-10-05
[Abstract]  Interactions between Hsp90, its co-chaperone Cdc37 and kinases have been biochemically studied for over three decades and have been shown to be functionally important in organisms from yeast to humans. However, formation of a stable complex for structural studies has been elusive. In this protocol we describe expression and purification of Hsp90-Cdc37-Cdk4 kinase protein complex from Saccharomyces cerevisiae utilizing the viral 2A sequences to titrate the three proteins at similar levels. [摘要]  Hsp90,其伴侣伴侣Cdc37和激酶之间的相互作用已经在三十多年的生物化学研究中被证明在酵母与人类的生物体内在功能上是重要的。 然而,形成一个稳定的结构研究复合物是难以捉摸的。 在该方案中,我们描述了利用病毒2A序列以相似水平滴定三种蛋白质的来自酿酒酵母的Hsp90-Cdc37-Cdk4激酶蛋白复合物的表达和纯化。
【背景】Hsp90分子伴侣与其客体激酶之间的稳定形成复合物已经被证明在体外是难治性的。以前的工作表明,Hsp90的共伴伴Cdc37与昆虫Sf9细胞中的客体激酶的过表达导致Sf9 Hsp90,外源Cdc37和外源激酶(Vaughan等人,2006)之间的稳定复合物。然而,昆虫细胞培养需要特殊的设备,比其他研究较好的表达系统(如细菌和酵母)要难以进行遗传操作,并且显着较慢地生长和克隆。上述蛋白质在E中的共表达。大肠杆菌不产生可溶性激酶/稳定复合物。我们认为,酿酒酵母将具有必要的机制来帮助折叠和促进复合物的形成,并试图通过共同表达这些蛋白质来产生人Hsp90β,人Cdc37和人Cdk4激酶之间的复合物, S上。酵母。为了获得三种蛋白质的化学计量表达,我们利用病毒2A肽,其允许三个蛋白质在一个mRNA上转录,随后在翻译阶段切割。该系统已被用于人类细胞系和兔网状细胞(Kim等人,2011; ...

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