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MicroAmpTM Optical Adhesive Film

MicroAmp ®光学胶粘膜

Company: Thermo Fisher Scientific
Catalog#: 4311971
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Characterizing the Transcriptional Effects of Endolysin Treatment on Established Biofilms of Staphylococcus aureus
Author:
Date:
2018-06-20
[Abstract]  Biofilms are the most common lifestyle of bacteria in both natural and human environments. The organized structure of these multicellular communities generally protects bacterial cells from external challenges, thereby enhancing their ability to survive treatment with antibiotics or disinfectants. For this reason, the search for new antibiofilm strategies is an active field of study. In this context, bacteriophages (viruses that infect bacteria) and their derived proteins have been proposed as promising alternatives for eliminating biofilms. For instance, endolysins can degrade peptidoglycan and, ultimately, lyse the target bacterial cells. However, it is important to characterize the responses of bacterial cells exposed to these compounds in order to improve the design of phage-based ... [摘要]  生物膜是自然和人类环境中最常见的细菌生活方式。这些多细胞社区的有组织结构通常保护细菌细胞免受外部挑战,从而增强其抗生素或消毒剂治疗的生存能力。为此,寻找新的抗菌膜策略是一个积极的研究领域。在这种情况下,已提出噬菌体(感染细菌的病毒)及其衍生蛋白作为消除生物膜的有希望的替代物。例如,内溶素可降解肽聚糖,并最终裂解靶细菌细胞。然而,表征暴露于这些化合物的细菌细胞的反应以改进基于噬菌体的抗微生物策略的设计是重要的。

如以前在Fernández等人(2017)中所描述的,开发该协议以检查暴露于内溶素处理的金黄色葡萄球菌生物膜细胞的转录反应。然而,它可能随后适用于分析其他微生物对不同抗菌剂的反应。

【背景】越来越清楚的是,亚抑制剂量的抗菌剂可能对目标微生物的不同表型具有调节作用,包括生物膜形成,代谢或毒力。因此,研究新化合物对低浓度靶细胞的潜在影响应该是发展过程的一部分。事实上,引发毒力因子或抗生素耐药决定簇产生的非常有效的抗菌剂可能不是治疗应用的良好候选者。另一方面,考虑到生物膜和浮游细胞之间的生理差异,应该对生物膜形成细胞分析新抗生物膜剂的作用似乎是合乎逻辑的。在这里,我们描述了一种协议,用于分析生物膜细胞在亚抑制浓度的内抑素浓度下的转录反应,噬菌体来源的蛋白质作为生物膜去除剂展现出巨大的前景。因此,通过RNA-seq将内溶素处理的细胞的转录组与对照细胞进行比较,并且后来通过RT-qPCR证实了所选基因的差异表达。 ...

Detection and Analysis of Circular RNAs by RT-PCR
Author:
Date:
2018-03-20
[Abstract]  Gene expression in eukaryotic cells is tightly regulated at the transcriptional and posttranscriptional levels. Posttranscriptional processes, including pre-mRNA splicing, mRNA export, mRNA turnover, and mRNA translation, are controlled by RNA-binding proteins (RBPs) and noncoding (nc)RNAs. The vast family of ncRNAs comprises diverse regulatory RNAs, such as microRNAs and long noncoding (lnc)RNAs, but also the poorly explored class of circular (circ)RNAs. Although first discovered more than three decades ago by electron microscopy, only the advent of high-throughput RNA-sequencing (RNA-seq) and the development of innovative bioinformatic pipelines have begun to allow the systematic identification of circRNAs (Szabo and Salzman, 2016; Panda et al., 2017b; Panda et al., ... [摘要]  真核细胞中的基因表达在转录和转录后水平受到严格调控。 mRNA转录,mRNA转录和mRNA翻译等后转录过程由RNA结合蛋白(RBPs)和非编码(nc)RNAs控制。大量的ncRNA家族包含多种调控RNA,如microRNAs和长的非编码(lnc)RNAs,但也是探索不足的一类环状RNAs。虽然三十多年前电子显微镜首次发现,但只有高通量RNA测序(RNA-seq)的出现和创新生物信息学管道的开发已经开始允许系统鉴定circRNA(Szabo和Salzman,2016;熊猫,2017b;熊猫等,2017c)。然而,通过RNA测序鉴定的真正的circRNA的验证需要其他分子生物学技术,包括常规或定量(q)聚合酶链反应(PCR)和Northern印迹分析(Jeck和Sharpless,2014)的逆转录(RT)。使用不同引物的环状RNA的RT-qPCR分析已被广泛用于检测,验证和有时定量circRNA(Abdelmohsen等人,2015和2017; Panda等人, ,2017b)。如在此详述的,设计为跨越循环RNA后接连接序列的分歧引物可以特异性扩增circRNA而不是对应的线性RNA。总之,使用不同引物的RT-PCR分析允许直接检测和定量circRNA。

【背景】CircRNAs是共价闭合的,缺少5'或3'末端的单链RNA。虽然它们的起源知之甚少,但它们可以通过称为反向剪接的过程从前体mRNA产生(Panda等人,2017d; ...

Polysome Fractionation to Analyze mRNA Distribution Profiles
Author:
Date:
2017-02-05
[Abstract]  Eukaryotic cells adapt to changes in external or internal signals by precisely modulating the expression of specific gene products. The expression of protein-coding genes is controlled at the transcriptional and post-transcriptional levels. Among the latter steps, the regulation of translation is particularly important in cellular processes that require rapid changes in protein expression patterns. The translational efficiency of mRNAs is altered by RNA-binding proteins (RBPs) and noncoding (nc)RNAs such as microRNAs (Panda et al., 2014a and 2014b; Abdelmohsen et al., 2014). The impact of factors that regulate selective mRNA translation is a critical question in RNA biology. Polyribosome (polysome) fractionation analysis is a powerful method to assess the association of ... [摘要]  真核细胞通过精确调节特定基因产物的表达来适应外部或内部信号的变化。蛋白质编码基因的表达受到转录和转录后水平的控制。在后面的步骤中,翻译的调节在需要蛋白质表达模式快速变化的细胞过程中特别重要。 mRNA的翻译效率由RNA结合蛋白(RBP)和非编码(nc)RNA如微RNA(Panda等人,2014a和2014b; Abdelmohsen等人)改变2014)。调节选择性mRNA翻译的因素的影响是RNA生物学中的一个关键问题。多核糖体(多核糖体)分馏分析是评估核糖体与给定mRNA的关联的有效方法。它提供了关于该mRNA的翻译状态的有价值的信息,这取决于与它们相关联的核糖体的数目,并且鉴定未翻译的mRNA(Panda等人,2016)。与许多核糖体相关的mRNA形成大量的多核糖体,预计将被主动翻译,而与少数或没有核糖体相关的mRNA有可能翻译不佳。总之,多聚糖分馏分析允许直接测定整个转录组水平的翻译效率以及个体mRNA。

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