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5'-二磷酸化寡核糖核苷酸的固相合成及其转化为帽化的m7Gppp-寡核糖核苷酸,用作甲型流感病毒RNA聚合酶体外引物。

Solid phase synthesis of 5'-diphosphorylated oligoribonucleotides and their conversion to capped m7Gppp-oligoribonucleotides for use as primers for influenza A virus RNA polymerase in vitro.

作者信息

Brownlee G G, Fodor E, Pritlove D C, Gould K G, Dalluge J J

机构信息

Chemical Pathology Unit, Sir William Dunn School of Pathology, University of Oxford, UK.

出版信息

Nucleic Acids Res. 1995 Jul 25;23(14):2641-7. doi: 10.1093/nar/23.14.2641.

Abstract

We have synthesized four different 5'-diphosphorylated oligoribonucleotides, varying in length from 11 to 13 nucleotides by a new solid phase method. After deprotection and partial purification the 5'-diphosphorylated oligoribonucleotides could be converted to capped (m7Gppp) oligoribonucleotides using guanylyl transferase. Radiolabelled capped oligoribonucleotides acted as primers for the influenza A virus RNA polymerase in vitro. The solid phase method described here should also allow the addition of 5'-diphosphates to synthetic oligodeoxyribonucleotides and be capable automation.

摘要

我们通过一种新的固相方法合成了四种不同的5'-二磷酸化寡核糖核苷酸,长度从11到13个核苷酸不等。脱保护和部分纯化后,使用鸟苷酰转移酶可将5'-二磷酸化寡核糖核苷酸转化为加帽(m7Gppp)寡核糖核苷酸。放射性标记的加帽寡核糖核苷酸在体外可作为甲型流感病毒RNA聚合酶的引物。本文所述的固相方法也应能将5'-二磷酸添加到合成寡脱氧核糖核苷酸上,并且能够实现自动化。

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