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UltraPureTM DNase/RNase-Free Distilled Water

UltraPure TM DNase / RNase-Free蒸馏水

Company: Thermo Fisher Scientific
Catalog#: 10977015
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Long-term in vitro Culture of Cryptosporidium parvum
Author:
Date:
2018-08-05
[Abstract]  Continuous in vitro growth of Cryptosporidium parvum has proved difficult and conventional in vitro culture techniques result in short-term (2-5 days) growth of the parasite resulting in thin-walled oocysts that fail to propagate using in vitro cultures, and do not produce an active infection using immunosuppressed or immunodeficient mouse models (Arrowood, 2002). Here we describe the use of hollow fiber bioreactors (HFB) that simulate in vivo conditions by providing oxygen and nutrients to host intestinal cells from the basal surface and permit the establishment of a low redox, high nutrient environment on the apical surface. When inoculated with 105 C. parvum (Iowa isolate) oocysts the bioreactor produced 108 ... [摘要]  Cryptosporidium parvum 的连续体外生长已证明是困难的,并且常规体外培养技术导致短期(2-5天)生长寄生虫导致薄壁卵囊不能使用体外培养物繁殖,并且不使用免疫抑制或免疫缺陷小鼠模型产生活跃感染(Arrowood,2002)。在这里,我们描述了中空纤维生物反应器(HFB)的使用,通过提供氧气和营养物质从基础表面宿主肠细胞模拟体内条件,并允许建立低氧化还原,高营养环境顶面。当接种10 5 C时。 parvum (爱荷华州分离物)卵囊生物反应器在14天后每ml产生10个 8 卵囊(20ml额外毛细血管体积),并保持2年以上。使用TCR-α免疫缺陷小鼠模型的体内感染性研究显示,在6,12和18个月时从生物反应器产生的卵囊与用于启动培养的亲本Iowa分离物无法区分。 HFB产生的卵囊具有与亲本爱荷华分离物类似的百分比分析。

【背景】 Cryptosporidium parvum 是人和其他哺乳动物肠道的细胞内专性寄生虫,导致急性腹泻。该疾病在免疫功能正常的个体中是自限性的,然而,在免疫功能低下的成人和幼儿中,该疾病可能危及生命(Kotloff,2017)。它是经济资源低的国家中三种被诊断出的儿童肠道疾病之一(Kotloff et al。,2013; Sow et ...

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 ...

Reduced Representation Bisulfite Sequencing in Maize
Author:
Date:
2018-03-20
[Abstract]  DNA methylation is an epigenetic modification that regulates plant development (Law and Jacobsen, 2010). Whole genome bisulfite sequencing (WGBS) is a state-of-the-art method for profiling genome-wide methylation patterns with single-base resolution (Cokus et al., 2008). However, for an organism with a large genome, e.g., the 2.1 Gb genome of maize, WGBS may be very expensive. Reduced representation bisulfite sequencing (RRBS) has been developed in mammalian studies (Smith et al., 2009). By digesting the genome with MspI with a size selection range of approximately 40-220 bp, CG-rich regions covering only ~1% of the human genome can be specifically sequenced. However, unlike mammalian genomes, plant genomes do not exhibit clear CpG islands. Therefore ... [摘要]  DNA甲基化是调节植物发育的表观遗传修饰(Law and Jacobsen,2010)。全基因组亚硫酸氢盐测序(WGBS)是用单碱基分辨率分析全基因组甲基化模式的最先进的方法(Cokus et al。,2008)。然而,对于具有大基因组的生物体,例如玉米的2.1Gb基因组,WGBS可能非常昂贵。代表性亚硫酸氢盐测序(RRBS)已经在哺乳动物研究中发展(Smith等人,2009)。通过用大小选择范围大约40-220bp的 Msp 消化基因组,可以对仅涵盖〜1%人类基因组的CG富含区域进行特异性测序。然而,与哺乳动物基因组不同,植物基因组不显示清楚的CpG岛。因此原来的RRBS协议不适用于工厂。因此,我们开发了一种计算机管道来选择特定的酶以生成感兴趣区域(ROI) - 富集的,例如,富含启动子的,减少的植物表达基因组(例如, Hsu et al。,2017)。通过用MseI消化玉米基因组并选择40-300bp片段,我们测序了大约四分之一的玉米基因组,同时保留了84.3%的启动子信息。该协议已在玉米中成功建立,可广泛应用于任何基因组。我们的计算机管道系统与RRBS文库制备方案相结合,允许进行计算分析和实验验证。

【背景】DNA甲基化是一种可遗传的表观遗传修饰,通过调节基因表达和染色质结构在动物,植物和真菌的许多发育过程中发挥重要作用(Law and ...

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