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MaxQTM 6000 Incubated/Refrigerated Stackable Shakers

MaxQ TM 6000孵化/冷藏可堆叠振荡器

Company: Thermo Fisher Scientific
Catalog#: SHKE6000
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EmPC-seq: Accurate RNA-sequencing and Bioinformatics Platform to Map RNA Polymerases and Remove Background Error
Author:
Date:
2021-02-20
[Abstract]  

Transcription errors can substantially affect metabolic processes in organisms by altering the epigenome and causing misincorporations in mRNA, which is translated into aberrant mutant proteins. Moreover, within eukaryotic genomes there are specific Transcription Error-Enriched genomic Loci (TEELs) which are transcribed by RNA polymerases with significantly higher error rates and hypothesized to have implications in cancer, aging, and diseases such as Down syndrome and Alzheimer’s. Therefore, research into transcription errors is of growing importance within the field of genetics. Nevertheless, methodological barriers limit the progress in accurately identifying transcription errors. Pro-Seq and NET-Seq can purify nascent RNA and map RNA polymerases along the genome but cannot be

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[摘要]  [摘要]转录错误可通过改变表观基因组并引起mRNA的错误整合而严重影响生物体内的代谢过程,从而将其翻译为异常的突变蛋白。此外,真核基因组内有特定转录错误富集的基因组基因座(TEELs),它们由RNA聚合酶与显著更高的错误率转录并推测为具有影响在癌症,老化和疾病例如唐氏综合征和阿尔茨海默'秒。因此,在遗传学领域对转录错误的研究越来越重要。尽管如此,方法上的障碍限制了准确识别转录错误的进展。Pro-Seq和NET-Seq可以沿基因组纯化新生RNA并绘制RNA聚合酶,但不能用于鉴定转录突变。在这里,我们本背景误差模型耦合的精密核圆形测序上运行(EMPC -SEQ),一种方法COMBIN荷兰国际集团测定和圆形测序核上运行与背景误差模型精确地检测新生转录错误和有效地辨别TEELs基因组中。

[背景]核糖核苷酸错掺导致的转录错误在所有活生物体中无处不在(Carey,2015)。假设每个信使RNA(mRNA)可以翻译2-4千次(Schwanhausser et al。,2011),并且许多特殊RNA在给定时间每个细胞仅表达一次(Islam et al。,2011; Pelechano et al。,2011)。,2010),即使是关键残基的单个转录错误也会使特定蛋白质的表达产生很大差异。另外,转录错误可加速蛋白质聚集,导致人类中与年龄有关的疾病(van ...

Heterologous Expression and Purification of Catalytic Domain of CESA1 from Arabidopsis thaliana
Author:
Date:
2016-10-20
[Abstract]  Heterologous expression of plant cellulose synthase (CESA) and its purification has remained a challenge for decades impeding detailed biophysical, biochemical and structural characterization of this key enzyme. An in-depth knowledge of structure and function of CESA proteins would enable us to better understand the hierarchical structure of the plant cell wall. Here, we report a detailed, and reproducible method of purification of catalytic domain of CESA1 from Arabidopsis thaliana that was recombinantly expressed in Escherichia coli. The method relies on a two stage purification procedure to obtain the catalytic domain in monomer and trimer forms. The biochemical and biophysical data including low resolution structures of the protein have been published (Vandavasi et ... [摘要]  植物纤维素合酶(CESA)的异源表达及其纯化在数十年来一直是阻碍该关键酶的详细生物物理,生物化学和结构表征的挑战。对CESA蛋白的结构和功能的深入了解将使我们能够更好地理解植物细胞壁的层次结构。在这里,我们报告了在大肠杆菌中重组表达的来自拟南芥的CESA1的催化结构域的详细的和可重复的纯化方法。该方法依赖于两阶段纯化方法以获得单体和三聚体形式的催化结构域。已经公开了包括蛋白质的低分辨率结构的生物化学和生物物理数据(Vandavasi等人,2016)。目前,这种蛋白质的结晶研究正在进行中。

[Backg 圆形] 纤维素是植物细胞壁最重要的结构组分,地球最大的生物可再生材料来源,但它的植物合成机制了解很少。植物纤维素合成复合物(CSC),也称为"玫瑰花结",因为其在电子显微镜图像中的六聚体外观,是一个大的多亚基跨膜蛋白复合物负责纤维素链的合成和它们组装成微原纤维。 ...

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