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RNasin® Ribonuclease Inhibitors

RNasin ® Plus RNA酶抑制剂

Company: Promega
Catalog#: N2611
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Host-regulated Hepatitis B Virus Capsid Assembly in a Mammalian Cell-free System
Author:
Date:
2018-04-20
[Abstract]  The hepatitis B virus (HBV) is an important global human pathogen and represents a major cause of hepatitis, liver cirrhosis and liver cancer. The HBV capsid is composed of multiple copies of a single viral protein, the capsid or core protein (HBc), plays multiple roles in the viral life cycle, and has emerged recently as a major target for developing antiviral therapies against HBV infection. Although several systems have been developed to study HBV capsid assembly, including heterologous overexpression systems like bacteria and insect cells, in vitro assembly using purified protein, and mammalian cell culture systems, the requirement for non-physiological concentrations of HBc and salts and the difficulty in manipulating host regulators of assembly presents major limitations ... [摘要]  乙型肝炎病毒(HBV)是一种重要的全球人类病原体,并且是肝炎,肝硬化和肝癌的主要原因。 HBV衣壳由单个病毒蛋白的多个拷贝组成,衣壳或核心蛋白(HBc)在病毒生命周期中起着多重作用,并且最近已经成为开发抗HBV病毒疗法的主要靶标。尽管已经开发了几种用于研究HBV衣壳组装的系统,包括异源过表达系统如细菌和昆虫细胞,使用纯化蛋白质和哺乳动物细胞培养系统进行体外组装,但对非生理浓度HBc和盐以及难以操纵装配的宿主调节物在生理相关条件下的衣壳装配的详细研究方面存在主要限制。我们最近开发了基于兔网织红细胞裂解物(RRL)的哺乳动物无细胞系统,其中HBc以生理浓度表达并在近生理条件下组装成衣壳。该系统已经揭示了HBc装配要求,这是以前装配系统所不能预料的。此外,该系统中的衣壳组装受可容易操作的内源宿主因子调控。在这里,我们提供了这种无细胞衣壳装配系统的详细协议,包括如何操纵调节装配的宿主因子的说明。

【背景】乙型肝炎病毒(HBV)是一种重要的全球人类病原体,其长期感染全世界数以亿计的人并且代表病毒性肝炎,肝硬化和肝癌的主要原因(Seeger等人, 2013; Trepo et。,2014)。 HBV通过逆转录RNA中间体(所谓的前基因组RNA(pgRNA))在核衣壳内(NC)复制其基因组DNA(一种宽松的环状部分双链DNA(RC ...

Adapting the Smart-seq2 Protocol for Robust Single Worm RNA-seq
Author:
Date:
2018-02-20
[Abstract]  Most nematodes are small worms that lack enough RNA for regular RNA-seq protocols without pooling hundred to thousand of individuals. We have adapted the Smart-seq2 protocol in order to sequence the transcriptome of an individual worm. While developed for individual Steinernema carpocapsae and Caenorhabditis elegans larvae as well as embryos, the protocol should be adaptable for other nematode species and small invertebrates. In addition, we describe how to analyze the RNA-seq results using the Galaxy online environment. We expect that this method will be useful for the studying gene expression variances of individual nematodes in wild type and mutant backgrounds. [摘要]  大多数线虫是小蠕虫,缺乏足够的RNA用于常规的RNA-seq协议,而没有汇集成千上万的个体。 我们已经调整了Smart-seq2协议来排序单个蠕虫的转录组。 虽然针对Steinernema carpocapsae和Caenorhabditis elegans幼虫以及胚胎开发,但该方案应该适用于其他线虫物种和小无脊椎动物。 另外,我们介绍如何使用Galaxy在线环境分析RNA-seq结果。 我们预计这种方法将有助于研究野生型和突变体背景个体线虫的基因表达差异。

【背景】低输入RNA-seq方案和扩增试剂盒,例如Smart-seq(Takara Bio,USA,Inc)和SuperAmp(Miltenyl Biotec,Inc),已经越来越多地开发和商业化,作为对低输入RNA-基于小组织,单一微生物和单细胞的seq研究。这些研究经常探索并解决特定群体(例如细胞群体,复杂组织或微生物群体)的个体中的异源基因表达。针对微生物(如线虫)的低输入RNA-seq方案的改进和适应将通过允许在单一线虫水平上分析基因表达异质性而极大地有益于线虫领域。在这里,我们已经调整了单细胞RNA-seq方案Smart-seq2(Picelli等人,2013和2014; Trombetta等人,2014),对于单线虫RNA测序。我们成功地在昆虫寄生线虫Steinernema ...

Immunoprecipitation of Tri-methylated Capped RNA
Author:
Date:
2018-02-05
[Abstract]  Cellular quiescence (also known as G0 arrest) is characterized by reduced DNA replication, increased autophagy, and increased expression of cyclin-dependent kinase p27Kip1. Quiescence is essential for wound healing, organ regeneration, and preventing neoplasia. Previous findings indicate that microRNAs (miRNAs) play an important role in regulating cellular quiescence. Our recent publication demonstrated the existence of an alternative miRNA biogenesis pathway in primary human foreskin fibroblast (HFF) cells during quiescence. Indeed, we have identified a group of pri-miRNAs (whose mature miRNAs were found induced during quiescence) modified with a 2,2,7-trimethylguanosine (TMG)-cap by the trimethylguanosine synthase 1 (TGS1) protein and transported to the cytoplasm ... [摘要]  蜂窝静止(因此已知为G <子> 0 骤停)是由降低的DNA复制,增加自噬表征,并且增加的细胞周期蛋白依赖性激酶p27蛋白上标kip1 表达。静止对伤口愈合,器官再生和瘤形成是必不可少的。先前的发现表明微小RNA(miRNA)在调节细胞静止过程中起重要作用。我们最近的出版物静止期间以实例阐述在原代人替代miRNA生物途径包皮成纤维(HFF)细胞的存在。实际上,我们已经发现了一组与由trimethylguanosine合酶1(TGS1)蛋白的2,2,7- trimethylguanosine(TMG)带肩改性PRI-的miRNA(其成熟miRNA发现静止期期间诱导的)的并运输到细胞质通过Exportin-1(XPO1)蛋白质。我们用来抗体针对(TMG)兴趣盖(不与第(m交叉反应 7 G)兴趣帽了大部分PRI-的miRNA或mRNA的含有[鲁曼等人的,1982]),以从增殖或静态HFFS的总RNA提取RNA进行免疫沉淀。该测定的新颖性是PRI-miRNA的以及含有一个TMG-帽修改以外的非编码RNA的特异性分离。


【背景】蜂窝静止,类型可逆生长停滞的,是在伤口愈合,器官再生,和预防瘤形成涉及一种重要的细胞状态(科勒,2011;瓦尔古等人 2012)。已发现小的非编码RNA如miRNA参与细胞静止的调节。 ...

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