Modified DNA bearing 5-(methoxycarbonyl-methyl)-2′-deoxyuridine:: Preparation by PCR with thermophilic DNA polymerase and post-synthetic derivation

被引：24

作者：

Masud, MM ^{[1
]}

Ozaki-Nakamura, A ^{[1
]}

Kuwahara, M ^{[1
]}

Ozaki, H ^{[1
]}

Sawai, H ^{[1
]}

机构：

[1] Gunma Univ, Dept Appl Chem, Gunma 3768515, Japan

来源：

CHEMBIOCHEM | 2003年 / 4卷 / 07期

关键词：

DNA modification; DNA polymerase; polymerase chain reaction; thymidine analogues; MODIFIED OLIGONUCLEOTIDES; HYBRIDIZATION PROPERTIES; CATALYTIC REPERTOIRE; THYMIDINE ANALOGS; NUCLEIC-ACIDS; NUCLEOTIDE; SELECTION; AMINO;

D O I：

10.1002/cbic.200200539

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A thymidine analogue bearing a methyl ester at the C5 position was accepted as a substrate by the thermophilic family B DNA polymerases, KOD Dash, Pwo, and Vent<LF>(exo-), to form the corresponding PCR product, but not by the thermophilic family A DNA polymerases, Taq, Tth, and T7 thermosequenase. Modified DNA containing this analogue was prepared by PCR on a large scale with KOD Dash DNA polymerase and 5-(methoxycarbonyl-methyl)-2'-deoxyuridine 5'-triphospate as a substrate. The methyl ester of the modified DNA was further allowed to react with tris (2-aminoethyl)amine or histamine by an ester-amide exchange reaction to form the corresponding derivatized DNA bearing a tris(2-aminoethyl)amine or histamine moiety. Hydrolysis of the methyl ester of the modified DNA gave a functionalized DNA bearing an anionic carboxyl group. The derivatized DNA could act as a template for the PCR with KOD Dash DNA polymerase and the natural 2'-deoxythymidine 5'-triphosphate or the modified thymidine analogue as a substrate. The postsynthetic derivatization of the modified DNA may expand the variety of structurally modified DNA produced by PCR.

引用

页码：584 / 588

页数：5

共 20 条

[1] Quantitative analysis of receptors for adenosine nucleotides obtained via in vitro selection from a library incorporating a cationic nucleotide analog [J].