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S equence‐specific M ethyltransferase‐ I nduced L abel ing of DNA (SMILing DNA)
Author(s) -
Pljevaljčić Goran,
Schmidt Falk,
Weinhold Elmar
Publication year - 2004
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200300739
Subject(s) - dna , methyltransferase , biology , dna methyltransferase , biochemistry , dna ligase , dna sequencing , microbiology and biotechnology , methylation
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. Taq I) 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.