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0.22 μm filter for small volumes

Company: Millipore
Catalog#: SLGV033RS
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Live Cell Imaging of Male Meiosis in Arabidopsis by a Landmark-based System
Author:
Date:
2020-05-05
[Abstract]  Live cell imaging has tremendously promoted our understanding of cellular and subcellular processes such as cell division. Here, we present a step-by-step protocol for a robust and easy-to-use live cell imaging approach to study male meiosis in the plant Arabidopsis thaliana as recently established. Our method relies on the concomitant analysis of two reporter genes that highlight chromosome configurations and microtubule dynamics. In combination, these reporter genes allowed the discrimination of five cellular parameters: cell shape, microtubule array, nucleus position, nucleolus position, and chromatin condensation. These parameters can adopt different states, e.g., the nucleus position can be central or lateral. Analyzing how tightly these states are associated gives ... [摘要]  [摘要 ] 活细胞成像极大地促进了我们对细胞分裂等亚细胞过程的了解。在此,我们提出了一种循序渐进的方案,以一种可靠且易于使用的活细胞成像方法来研究雄性减数分裂。植物拟南芥。由于最近建立了我们的方法依赖于伴随分析中的两个报告基因,突出染色体构和微管动态组合,这些记者基因准许入住歧视五细胞参数:细胞形状,微管阵列,核POSITI 在,核仁位置,染色质凝聚,这些参数可以采用不同的国家,例如,该核位置可以是中央或外侧。分析如何紧密这些国家有关联产生了里程碑阶段,反过来允许定量和定性的解剖减数分裂过程我们设想,这种方法还可以为细胞分化分析提供有价值的标准 减数分裂之外的增强过程。

[背景 ] 减数分裂是一种特殊的细胞分裂周期,有两个主要目的:首先,减数分裂母细胞的DNA含量降低一半,在二倍体生物体中会导致单倍体减数分裂产物。有性繁殖的生物,以便在受精过程中融合了两个配子后,恢复了原始的基因组大小。其次,减数分裂通过亲本染色体(同源染色体或短同源)之间的DNA片段交换(称为减数分裂重组)促进了遗传多样性。通过生成新的但完整的染色体集,其中每个染色体随机存在母亲或父亲的同系物(如果是二倍体生物体)。因此,减数分裂对于不同领域的研究很有趣,从细胞生物学和生殖生物学,从遗传学到进化生物学(Wijn ker 和Schnittger ,2013; Mercier ...

Preparation of a Bacteriophage T4-based Prokaryotic-eukaryotic Hybrid Viral Vector for Delivery of Large Cargos of Genes and Proteins into Human Cells
Author:
Date:
2020-04-05
[Abstract]  A viral vector that can safely and efficiently deliver large and diverse molecular cargos into cells is the holy grail of curing many human diseases. Adeno-associated virus (AAV) has been extensively used but has a very small capacity. The prokaryotic virus T4 has a large capacity but lacks natural mechanisms to enter mammalian cells. Here, we created a hybrid vector by combining T4 and AAV into one nanoparticle that possesses the advantages of both. The small 25 nm AAV particles are attached to the large 120 nm x 86 nm T4 head through avidin-biotin cross-bridges using the phage decoration proteins Soc (small outer capsid protein) and Hoc (highly antigenic outer capsid protein). AAV thus “piggy-backed” on T4 capsid, by virtue of its natural ability to enter many types of human cells ... [摘要]  [摘要 ] 一种病毒载体,可以安全有效地将大量多样的分子货物运送到细胞中 是治愈许多人类疾病的圣杯。腺伴随病毒(AAV)已被广泛使用,但容量很小。T4原核病毒容量大,但缺乏进入哺乳动物细胞的天然机制。在这里,我们通过将T4和AAV结合到一个具有两者优势的纳米颗粒中,创建了一种杂交载体。使用噬菌体修饰蛋白Soc(小的外衣壳蛋白)和Hoc(高度抗原化的外衣壳蛋白),通过亲和素-生物素交叉桥将25 nm的AAV小颗粒连接到120 nm x 86 nm的大T4头上。因此,AAV凭借其固有的进入多种类型人体细胞的自然能力,可以“背负”于T4衣壳上,从而有效地充当了“驱动器”,以运送与T4头相关的大型货物。这种独特的T4-AAV杂交载体方法可为将来开发新型疗法铺平道路。

[背景 ] 已经有新的和有效的递送载体能够运输基因和蛋白质的大货物进入人类细胞,以刺激生产治疗性生物分子的和/或修复的细胞和遗传缺陷的迫切需要。这样的载体将允许将快速出现的技术(例如CRISPR,CAR T细胞等)转化为用于大规模应用以及个性化医学的疗法(Stewart 等,2016)。

将具有不同特性的纳米粒子组装到杂化复合物中是开发新型功能材料的有力策略,因为这些杂化复合物显示出集体和协作的属性,其中某些属性可能与单个粒子所显示的属性不同(Ghosh 等人,2012; ...

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