用T4 RNA连接酶将单核苷酸添加到寡核糖核苷酸受体上。
Addition of mononucleotides to oligoribonucleotide acceptors with T4 RNA ligase.
作者信息
Kikuchi Y, Hishinuma F, Sakaguchi K
出版信息
Proc Natl Acad Sci U S A. 1978 Mar;75(3):1270-3. doi: 10.1073/pnas.75.3.1270.
Abstract
RNA ligase induced by bacteriophage T4 catalyzed the addition of nucleoside 5',3'-diphosphates to oligoribonucleotide acceptors in the presence of ATP. The reactions proceeded in equimolar concentrations of donors and acceptors. Several oligonucleotides of defined sequence were synthesized by this method in yields of 28-68%. The enzyme required the phosphate ester at the 3' position of the donor molecule, nucleoside 5',2'-diphosphates being unable to serve as donors. Thymidine 5',3'-diphosphate was active as a donor for the enzyme. The ligation product, (Ap)2ApCp, was also obtained from the reaction of (Ap)2A and 5'-adenylylated cytidine 5',3'-diphosphate (A5'pp5'Cp) with RNA ligase in the absence of ATP. These results show that the minimal substrate for RNA ligase is a nucleoside 5',3'-diphosphate.
摘要
噬菌体T4诱导的RNA连接酶在ATP存在的情况下，催化将核苷5',3'-二磷酸添加到寡核糖核苷酸受体上。反应在供体和受体等摩尔浓度下进行。通过这种方法合成了几种特定序列的寡核苷酸，产率为28 - 68%。该酶需要供体分子3'位置的磷酸酯，核苷5',2'-二磷酸不能作为供体。胸苷5',3'-二磷酸作为该酶的供体具有活性。连接产物(Ap)2ApCp也可在无ATP的情况下，由(Ap)2A和5'-腺苷酸化的胞苷5',3'-二磷酸(A5'pp5'Cp)与RNA连接酶反应获得。这些结果表明，RNA连接酶的最小底物是核苷5',3'-二磷酸。
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