High-yield production of short GpppA- and 7MeGpppA-capped RNAs and HPLC-monitoring of methyltransfer reactions at the guanine-N7 and adenosine-2′O positions
High-yield production of short GpppA- and 7MeGpppA-capped RNAs and HPLC-monitoring of methyltransfer reactions at the guanine-N7 and adenosine-2′O positions
复制标题
DOI：
10.1093/nar/gkl1119
复制
发表时间：
2007-02-01
影响因子：
14.9
通讯作者：
Alvarez, K.
中科院分区：
文献类型：
--
作者：
Peyrane, F.;Selisko, B.;Alvarez, K.
许多真核生物和病毒的mRNA，其中第一个转录的核苷酸是腺苷，用cap-1结构，（7 Me）G（5 '）-ppp（5'）-A（2 'OMe）修饰。例如，黄病毒（登革热、西尼罗河病毒）的正义RNA基因组显示腺苷的严格保守性。我们着手生产GpppA-和（7 Me）GpppA-加帽的RNA寡核苷酸用于非放射性mRNA帽甲基转移酶测定，并在透视图中，用于与底物长度相关的酶特异性研究以及共结晶研究。本研究报告了使用噬菌体T7 DNA引发酶片段合成GpppAC（n）和（7 Me）GpppAC（n）（1
Many eukaryotic and viral mRNAs, in which the first transcribed nucleotide is an adenosine, are decorated with a cap-1 structure, (7Me)G(5')-ppp(5')-A(2'OMe). The positive-sense RNA genomes of flaviviruses (Dengue, West Nile virus) for example show strict conservation of the adenosine. We set out to produce GpppA- and (7Me)GpppA-capped RNA oligonucleotides for non-radioactive mRNA cap methyltransferase assays and, in perspective, for studies of enzyme specificity in relation to substrate length as well as for co-crystallization studies. This study reports the use of a bacteriophage T7 DNA primase fragment to synthesize GpppAC(n) and (7Me)GpppAC(n) (1