Bird Jeremy G, Nickels Bryce E, Ebright Richard H
Department of Genetics and Waksman Institute, Rutgers University, Piscataway, USA.
Department of Chemistry and Waksman Institute, Rutgers University, Piscataway, USA.
Bio Protoc. 2017 Jun 20;7(12). doi: 10.21769/BioProtoc.2336.
It recently has been established that adenine-containing cofactors, including nicotinamide adenine dinucleotide (NAD), reduced nicotinamide adenine dinucleotide (NADH), and 3'-desphospho-coenzyme A (dpCoA), can serve as 'non-canonical initiating nucleotides' (NCINs) for transcription initiation by bacterial and eukaryotic cellular RNA polymerases (RNAPs) and that the efficiency of the reaction is determined by promoter sequence (Bird , 2016). Here we describe a protocol to quantify the relative efficiencies of transcription initiation using an NCIN vs. transcription initiation using a nucleoside triphosphate (NTP) for a given promoter sequence.
最近已经确定，含腺嘌呤的辅因子，包括烟酰胺腺嘌呤二核苷酸（NAD）、还原型烟酰胺腺嘌呤二核苷酸（NADH）和3'-去磷酸辅酶A（dpCoA），可以作为细菌和真核细胞RNA聚合酶（RNAPs）转录起始的“非经典起始核苷酸”（NCINs），并且反应效率由启动子序列决定（伯德，2016年）。在这里，我们描述了一种协议，用于量化使用NCIN进行转录起始的相对效率与使用核苷三磷酸（NTP）进行转录起始的相对效率，以确定给定启动子序列。
