一种使用多核苷酸磷酸化酶通过单次添加2'-O-(邻硝基苄基)核苷5'-二磷酸来合成寡核苷酸的方法。

A method for the synthesis of oligonucleotide by single addition of 2'-O-(o-nitrobenzyl)nucleoside 5'-diphosphates using polynucleotide phosphorylase.

作者信息

Ohtsuka E, Tanaka S, Hayashi M, Ikehara M

出版信息

Biochim Biophys Acta. 1979 Nov 22;565(1):192-8. doi: 10.1016/0005-2787(79)90095-9.

DOI:10.1016/0005-2787(79)90095-9

Abstract

Two hexanucleotides A-U-G-U-G-A and C-A-A-U-U-G were synthesized from the chemically synthesized trimers C-A-A and A-U-G by addition of 2'-O-(o-nitrobenzyl)nucleoside diphosphates using polynucleotide phosphorylase isolated from either Escherichia coli or Micrococcus luteus. In each reaction the preference of the enzyme was tested. The o-nitrobenzyl group was removed after addition of the mononucleotide and the deblocked product was isolated by chromatography on DEAE-Sephadex in high yields.

摘要

通过使用从大肠杆菌或藤黄微球菌中分离得到的多核苷酸磷酸化酶，添加2'-O-(邻硝基苄基)核苷二磷酸，由化学合成的三聚体C-A-A和A-U-G合成了两个六核苷酸A-U-G-U-G-A和C-A-A-U-U-G。在每个反应中都测试了该酶的偏好性。在添加单核苷酸后去除邻硝基苄基，并通过在DEAE-葡聚糖凝胶上进行色谱分离，以高收率分离出脱保护产物。

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