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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.

DOI:10.1073/pnas.75.3.1270
PMID:274717
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC411452/
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。

相似文献

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Addition of mononucleotides to oligoribonucleotide acceptors with T4 RNA ligase.用T4 RNA连接酶将单核苷酸添加到寡核糖核苷酸受体上。
Proc Natl Acad Sci U S A. 1978 Mar;75(3):1270-3. doi: 10.1073/pnas.75.3.1270.
2
Enzymatic oligoribonucleotide synthesis with T4 RNA ligase.利用T4 RNA连接酶进行酶促寡核糖核苷酸合成。
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Equimolar addition of oligoribonucleotides with T4 RNA ligase.用T4 RNA连接酶等摩尔添加寡核糖核苷酸。
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[Mechanism of reaction catalyzed by RNA-ligase from bacteriophage T4].[噬菌体T4 RNA连接酶催化的反应机制]
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本文引用的文献

1
A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. I. Purification and characterization of alkaline phosphatase.大肠杆菌碱性磷酸酶的精细结构遗传学与化学研究。I. 碱性磷酸酶的纯化与特性分析
Biochim Biophys Acta. 1960 Mar 11;38:470-83. doi: 10.1016/0006-3002(60)91282-8.
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Phosphorylation of nucleic acid by an enzyme from T4 bacteriophage-infected Escherichia coli.来自T4噬菌体感染的大肠杆菌的一种酶对核酸的磷酸化作用。
Proc Natl Acad Sci U S A. 1965 Jul;54(1):158-65. doi: 10.1073/pnas.54.1.158.
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Monofunctional substrates of polynucleotide phosphorylase. The monoaddition of 2'(3')-O-isovaleryl-nucleoside diphosphate to an initiator oligonucleotide.多核苷酸磷酸化酶的单功能底物。2'（3'）-O-异戊酰核苷二磷酸向引发寡核苷酸的单加成。
Eur J Biochem. 1971 Dec 22;24(1):4-11. doi: 10.1111/j.1432-1033.1971.tb19649.x.
4
New approach to the synthesis of polyribonucleotides of defined sequence.合成特定序列多聚核糖核苷酸的新方法。
Nature. 1971 Oct 22;233(5321):551-3. doi: 10.1038/233551a0.
5
Studies on ribonucleic acid ligase. Characterization of an adenosine triphosphate-inorganic pyrophosphate exchange reaction and demonstration of an enzyme-adenylate complex with T4 bacteriophage-induced enzyme.核糖核酸连接酶的研究。三磷酸腺苷-无机焦磷酸交换反应的特性以及与T4噬菌体诱导酶形成的酶-腺苷酸复合物的证明。
J Biol Chem. 1974 Dec 10;249(23):7447-56.
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Purification and properties of bacteriophage T4-induced RNA ligase.噬菌体T4诱导的RNA连接酶的纯化及性质
Proc Natl Acad Sci U S A. 1972 Oct;69(10):3009-13. doi: 10.1073/pnas.69.10.3009.
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A method for rapid separation of [32P]-inorganic phosphate or pyrophosphate from ATP.
Anal Biochem. 1970 Dec;38(2):383-8. doi: 10.1016/0003-2697(70)90461-6.
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'Single addition' and 'transnucleotidation' reactions catalyzed by polynucleotide phosphorylase. Effect of enzymatic removal of inorganic phosphate during reaction.多核苷酸磷酸化酶催化的“单加成”和“转核苷酸作用”反应。反应过程中酶促去除无机磷酸盐的影响。
Nucleic Acids Res. 1974 Dec;1(12):1665-74. doi: 10.1093/nar/1.12.1665.
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Novel monofunctional substrates of polynucleotide phosphorylase. The "single-addition" of 2'(3')-O-dihydrocinnamoyl-nucleoside 5'-diphosphate to a primer oligonucleotide.多核苷酸磷酸化酶的新型单功能底物。2'（3'）-O-二氢肉桂酰核苷5'-二磷酸对引物寡核苷酸的“单次添加”。
J Biochem. 1975 Feb;77(2):469-72. doi: 10.1093/oxfordjournals.jbchem.a130747.
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Determination of recognition sites of T4 RNA ligase on the 3'-OH and 5' -P termini of polyribonucleotide chains.T4 RNA连接酶在多聚核糖核苷酸链3'-OH和5'-P末端识别位点的确定
Nature. 1975 Apr 3;254(5499):452-4. doi: 10.1038/254452a0.