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从寡核糖核苷酸磷酸三酯上去除核苷酸间的2,2,2-三氯乙基保护基团后，使用葡聚糖凝胶LH20进行色谱分离作为一种有效的纯化步骤。

Chromatography on Sephadex LH20 as an efficient purification step after removal of internucleotide 2,2,2-trichloroethyl protective groups from oligoribonucleotide phosphotriesters.

作者信息

Grześkowiak K, Adamiak R W, Wiewiórowski M

出版信息

Nucleic Acids Res. 1980 Mar 11;8(5):1097-105. doi: 10.1093/nar/8.5.1097.

DOI:10.1093/nar/8.5.1097
PMID:7443541
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC323975/
Abstract

Chromatography on Sephadex LH20, in a linear gradient of methanol in 0.02M TEAB buffer pH 7.5, is proposed as a fast and efficient method for the isolation and purification of protected oligoribonucleotide phosphodiesters obtained by deprotection of internucleotide phosphotriesters, and for the monitoring of the deprotection step itself. Its utility is shown on the example of removal of 2,2,2-trichloroethyl groups from oligoribonucleotide phosphotriester I of sequence CCCAUAA by two methods: /1/ reductive elimination with zinc in the presence of acetylacetone modified as presented here, and /2/ hydrogenolytic dehalogenation over palladium in pyridine. This method of chromatography on Sephadex LH20 is used as a key purification step during the removal of 2,2,2-trichloroethyl groups from I by method /1/ and allows to raise the yield of III during fianl deprotection step from 5 to 65%.

摘要

在0.02M pH 7.5的TEAB缓冲液中以甲醇线性梯度进行的Sephadex LH20柱色谱法，被提议作为一种快速有效的方法，用于分离和纯化通过核苷酸间磷酸三酯脱保护得到的受保护寡核糖核苷酸磷酸二酯，以及用于监测脱保护步骤本身。以通过两种方法从序列为CCCAUAA的寡核糖核苷酸磷酸三酯I中去除2,2,2-三氯乙基基团为例展示了其效用：/1/ 在此处所述的经乙酰丙酮修饰的情况下用锌进行还原消除，以及 /2/ 在吡啶中于钯上进行氢解脱卤。这种Sephadex LH20柱色谱法被用作通过方法 /1/ 从I中去除2,2,2-三氯乙基基团过程中的关键纯化步骤，并使得在最终脱保护步骤中III的产率从5%提高到65%。

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本文引用的文献

1
Sites and thermodynamic quantities associated with proton and metal ion interaction with ribonucleic acid, deoxyribonucleic acid, and their constituent bases, nucleosides, and nucleotides.与质子和金属离子与核糖核酸、脱氧核糖核酸及其组成碱基、核苷和核苷酸相互作用相关的位点和热力学量。
Chem Rev. 1971 Oct;71(5):439-81. doi: 10.1021/cr60273a002.
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Deoxy oligonucleotide synthesis via the triester method.通过三酯法合成脱氧寡核苷酸。
J Org Chem. 1973 Jan 26;38(2):245-50. doi: 10.1021/jo00942a011.
3
Studies on transfer ribonucleic acids and related compounds. VI. Synthesis of yeast alanine transfer ribonucleic acid 3'-terminal nonanucleotides and 5'-terminal hexanucleotides.转移核糖核酸及相关化合物的研究。VI. 酵母丙氨酸转移核糖核酸3'-末端九核苷酸和5'-末端六核苷酸的合成。
J Am Chem Soc. 1973 Jul 11;95(14):4725-33. doi: 10.1021/ja00795a042.
4
Nucleoside 3'-phosphotriesters as key intermediates for the oligoribonucleotide synthesis. IV. New method for removal of 2,2,2-trichloroethyl group and 31P NMR as a new tool for analysis of deblocking of internucleotide phosphate protecting groups.核苷3'-磷酸三酯作为寡核糖核苷酸合成的关键中间体。IV. 去除2,2,2-三氯乙基基团的新方法以及31P NMR作为分析核苷酸间磷酸保护基团脱保护的新工具。
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5
The chemical synthesis of the anticodon loop of an eukaryotic initiator tRNA containing the hypermodified nucleoside N6-/N-threonylcarbonyl/-adenosine/t6A/1.包含超修饰核苷N6-（N-苏氨甲酰）-腺苷（t6A）的真核起始tRNA反密码子环的化学合成。
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6
The phosphotriester approach to oligonucleotide synthesis: Preparation of oligo- and poly-thymidylic acids.用于寡核苷酸合成的磷酸三酯法：寡聚胸苷酸和聚胸苷酸的制备
J Chem Soc Perkin 1. 1977;4:445-60. doi: 10.1039/p19770000445.
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Neighbouring group participation in the unblocking of phosphotriesters of nucleic acids.相邻基团参与核酸磷酸三酯的解封闭反应。
Nucleic Acids Res. 1979;6(6):2237-59. doi: 10.1093/nar/6.6.2237.

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