{{'Search' | translate}}
 

Ethylene Glycol Bis(β-aminoethylether)-N,N,N',N'-tetraacetic Acid [EGTA;GEDTA] Nuclease and Protease tested

Company: NACALAI TESQUE
Catalog#: 08907-42
Bio-protocol()
Company-protocol()
Other protocol()

Time-lapse Observation of Chromosomes, Cytoskeletons and Cell Organelles during Male Meiotic Divisions in Drosophila
Author:
Date:
2017-04-20
[Abstract]  In this protocol, we provide an experimental procedure that perform time-lapse observation of intra-cellular structures such as chromosomes, cytoskeletons and cell organelles during meiotic cell divisions in Drosophila males. As primary spermatocyte is the largest dividing diploid cell in Drosophila, which is equivalent in size to mammalian cultured cells, one can observe dynamics of cellular components during division of the model cells more precisely. Using this protocol, we have showed that a microtubule-associated protein plays an essential role in microtubule dynamics and initiation of cleavage furrowing through interaction between microtubules and actomyosin filaments. We have also reported that nuclear membrane components are required for a formation and/or ... [摘要]  在这个协议中,我们提供了一个实验程序,在果蝇男性的减数分裂细胞分裂期间,进行细胞内结构如染色体,细胞骨架和细胞器的延时观察。由于主要精母细胞是果蝇中最大的分裂二倍体细胞,其大小与哺乳动物培养细胞相当,可以更准确地观察模型细胞分裂期间细胞组分的动力学。使用该方案,我们已经表明,微管相关蛋白在微管动力学中起重要作用,并通过微管和肌动球蛋白丝之间的相互作用开始裂解沟槽。我们还报道说,需要核膜组分来形成和/或维持在果蝇细胞中细胞分裂所必需的主轴包络。

在果蝇中,也可以在标准培养条件下良好培养的良好培养细胞系。然而,它们的单元尺寸,特别是细胞质体积,比哺乳动物细胞的小得多。这在细胞分裂过程中损害了细胞成分的检查。精母细胞,在另一方面,实现第一次减数分裂开始之前不同的细胞生长。主要精母细胞是出现在果蝇发育中的增殖细胞中最大的二倍体细胞。因此,可以使用光学显微镜容易地细分观察分裂细胞中的细胞结构。在果蝇黑腹果蝇中,提供了先进和复杂的遗传技术(Ashburner等人,2004)。染色体分离和细胞分裂中的减数分裂缺陷出现在完成2< 减数分裂后精子细胞的细胞组织中。通过观察这种早期精子细胞,人们可以很容易地发现甚至微小的减数分裂异常(2012); 2012年; ...

RNase Sensitivity Screening for Nuclear Bodies with RNA Scaffolds in Mammalian Cells
Author:
Date:
2017-04-20
[Abstract]  The mammalian cell nucleus is highly organized and contains membraneless nuclear bodies (NBs) characterized by distinct resident factors. The NBs are thought to serve as sites for biogenesis and storage of certain RNA and protein factors as well as assembly of ribonucleoprotein complexes. Some NBs are formed with architectural RNAs (arcRNAs) as their structural scaffolds and additional NBs likely remain unidentified in mammalian cells. Here, we describe an experimental protocol to search for new NBs built on certain arcRNAs. RNase-sensitive NBs were identified by monitoring nuclear foci visualized by tagging thousands of human cDNA products. [摘要]  哺乳动物细胞核高度组织,包含以不同的居民因素为特征的无膜核体(NBs)。 NB被认为是用于某些RNA和蛋白质因子的生物发生和储存的位点以及核糖核蛋白复合物的组装。 一些NB由构建的RNA(arcRNA)形成,作为它们的结构性支架,另外的NB可能在哺乳动物细胞中保持不明。 在这里,我们描述了一个实验协议来搜索建立在某些arcRNA上的新NB。 通过监测通过标记数千个人类cDNA产物可视化的核病灶来鉴定RNase敏感性NB。
【背景】哺乳动物细胞核是高度组织的,由称为核体(NB)的多个不同的亚结构组成。迄今为止,已经将15个NB鉴定为含有各种蛋白质和RNA因子的亚核膜无颗粒结构,其中许多颗粒结构用作特异性RNA,蛋白质和/或核糖核蛋白(RNP)复合物的生物发生,成熟,储存和螯合的位点Mao et al。,2011; Sleeman and Trinkle-Mulcahy,2014)(表1)。
一些NB被构建在称为结构RNA(arcRNA)的特定长非编码RNA(lncRNA)上,其定义为NB的结构核心(Chujo等,2016)。 arcRNA依赖的NB由与arcRNA相互作用的许多RNA结合蛋白组成。最显着的例子是由几种特征性RNA结合蛋白组成的寄生虫斑(Fox等,2002; ...

Comments