Sequence-specific methyltransferase-induced labeling of DNA (SMILing DNA)

被引：60

作者：

Pljevaljcic, G

Schmidt, F

Weinhold, E

机构：

[1] Rhein Westfal TH Aachen, Inst Organ Chem, D-52056 Aachen, Germany

[2] Scripps Res Inst, Dept Mol Biol, La Jolla, CA 92037 USA

来源：

CHEMBIOCHEM | 2004年 / 5卷 / 03期

关键词：

bioorganic chemistry; cofactors; DNA methylation; enzyme catalysis; transferases;

D O I：

10.1002/cbic.200300739

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 new concept for sequence-specific labeling of DNA by using chemically modified cofactors for DNA methyltransferases is presented. Replacement of the amino acid side chain of the natural cofactor S-adenosyl-L-methionine with an aziridine group leads to a cofactor suitable for DNA methyltransferase-catalyzed sequence-specific coupling with DNA. Sequence-specifically fluorescently labeled plasmid DNA was obtained by using the DNA methyltransferase from Thermus aquaticus (M.Taql) as catalyst and attaching a fluorophore to the aziridine cofactor. First results suggest that all classes of DNA methyltransferases with different recognition sequences can be used. In addition, this novel method for DNA labeling should be applicable to a wide variety of reporter groups.

引用

页码：265 / 269

页数：5

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