Author:
Date:
2018-08-05
[Abstract] This is a protocol for quantitative determination of storage and total carbohydrates in algae and cyanobacteria. The protocol is simple, fast and sensitive and it requires only few standard chemicals. Great advantage of this protocol is that both storage and total saccharides can be determined in the cellular pellets that were already used for chlorophyll and carotenoids quantification. Since it is recommended to perform the pigments measurement in triplicates, each pigment analysis can generate samples for both total saccharide and glycogen/starch content quantification.
The protocol was applied for quantification of both storage and total carbohydrates in cyanobacteria Synechocystis sp. PCC 6803, Cyanothece sp. ATCC 51142 and Cyanobacterium sp. ...
[摘要] 这是用于定量测定藻类和蓝细菌中的储存和总碳水化合物的方案。该协议简单,快速,灵敏,只需要很少的标准化学品。该方案的最大优点是可以在已经用于叶绿素和类胡萝卜素定量的细胞沉淀中测定储存和总糖。由于建议一式三份进行颜料测量,因此每种颜料分析都可以生成总糖和糖原/淀粉含量定量的样品。
该方案用于量化蓝细菌 Synechocystis sp中的储存和总碳水化合物。 PCC 6803, Cyanothece sp。 ATCC 51142和 Cyanobacterium sp。 IPPAS B-1200。它还被用于估算 Galdieria (IPPAS P-500,IPPAS P-507,IPPAS P-508,IPPAS P-513), Cyanidium caldarium IPPAS中的储存多糖P-510,绿藻小球藻 sp。 IPPAS C-1和C-1210, Parachlorella kessleri IPPAS C-9, Nannochloris sp。 C-1509, Coelastrella sp。 IPPAS H-626, Haematococcus sp。 IPPAS H-629和H-239,以及 Eustigmatos sp。 IPPAS H-242和IPPAS C-70。
【背景】碳水化合物在藻类和蓝细菌的代谢中起着许多作用。正如Raven和Beardall(Raven和Beardall,2003)所总结的那样,碳水化合物代表碳还原/氧化途径(光合作用和光呼吸)中间体的主要汇集,它们提供生长所需的骨架(例如,for氨基酸或细胞壁生物合成),它们代表ATP和还原当量的来源(通过呼吸途径),它们作为相容的溶质,它们对维持细胞膨胀是必不可少的(通过确保细胞壁的刚性结构)并且它们可以清除自由基。储存多糖(其中藻类和蓝藻中的主要形式是淀粉和糖原)作为能量和碳源,缓冲碳水化合物生产和消费率之间的不成比例,并允许活跃的新陈代谢(例如,固氮)在黑暗时期。 ...
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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; ...
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