Selenium modification of nucleic acids: preparation of phosphoroselenoate derivatives for crystallographic phasing of nucleic acid structures

被引：12

作者：

Pallan, Pradeep S. ^{[1
]}

Egli, Martin ^{[1
]}

机构：

[1] Vanderbilt Univ, Sch Med, Dept Biochem, Nashville, TN 37232 USA

来源：

NATURE PROTOCOLS | 2007年 / 2卷 / 03期

关键词：

X-RAY CRYSTALLOGRAPHY; HEAVY-ATOM DERIVATIVES; ENZYMATIC-SYNTHESIS; CRYSTAL-STRUCTURE; MAD; DNA; OLIGONUCLEOTIDES; RNA; 5'-(ALPHA-P-SELENO)TRIPHOSPHATE; OLIGORIBONUCLEOTIDES;

D O I：

10.1038/nprot.2007.74

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

This protocol describes a simplified means of introducing an anomalously scattering atom into oligonucleotides by conventional solid-phase synthesis. Replacement of a nonbridging phosphate oxygen in the backbone with selenium is practically suitable for any nucleic acid. The resulting oligonucleotide P-diastereomers can be separated using anion exchange HPLC to yield diastereomerically pure phosphoroselenoates (PSes). The total time for the synthesis and ion-exchange HPLC separation of pure PSe is approximately 60 h.

引用

页码：640 / 646

页数：7

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