School of Chemistry, The University of Nottingham, University Park, Nottingham, NG7 2RD, UK.
Sci Rep. 2011;1:126. doi: 10.1038/srep00126. Epub 2011 Oct 24.
A method is described for the detection of certain nucleotide modifications adjacent to the 5' 7-methyl guanosine cap of mRNAs from individual genes. The method quantitatively measures the relative abundance of 2'-O-methyl and N(6),2'-O-dimethyladenosine, two of the most common modifications. In order to identify and quantitatify the amounts of N(6),2'-O-dimethyladenosine, a novel method for the synthesis of modified adenosine phosphoramidites was developed. This method is a one step synthesis and the product can directly be used for the production of N(6),2'-O-dimethyladenosine containing RNA oligonucleotides. The nature of the cap-adjacent nucleotides were shown to be characteristic for mRNAs from individual genes transcribed in liver and testis.
介绍了一种用于检测特定核苷酸修饰的方法，这些修饰位于来自单个基因的 mRNA 的 5' 7-甲基鸟苷帽的附近。该方法定量测量了两种最常见的修饰物，即 2'-O-甲基和 N(6),2'-O-二甲基腺苷的相对丰度。为了鉴定和定量 N(6),2'-O-二甲基腺苷的含量，开发了一种用于合成修饰腺苷磷酰胺的新方法。该方法是一步合成，产物可直接用于合成含有 N(6),2'-O-二甲基腺苷的 RNA 寡核苷酸。结果表明，肝和睾丸中单个基因转录的 mRNA 帽附近核苷酸的性质具有特征性。
