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As Fast and Selective as Enzymatic Ligations: Unpaired Nucleobases Increase the Selectivity of DNA‐Controlled Native Chemical PNA Ligation
Author(s) -
Ficht Simon,
Dose Christian,
Seitz Oliver
Publication year - 2005
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.200500229
Subject(s) - native chemical ligation , dna ligase , sequencing by ligation , ligation , oligonucleotide , dna , chemistry , chemical ligation , ap site , nucleic acid , combinatorial chemistry , biochemistry , biology , dna damage , microbiology and biotechnology , enzyme , cysteine , genomic library , base sequence
Abstract DNA‐controlled reactions offer interesting opportunities in biological, chemical, and nanosciences. In practical applications, such as in DNA sequence analysis, the sequence fidelity of the chemical‐ligation reaction is of central importance. We present a ligation reaction that is as fast as and much more selective than enzymatic T4 ligase‐mediated oligonucleotide ligations. The selectivity was higher than 3000‐fold in discriminating matched from singly mismatched DNA templates. It is demonstrated that this enormous selectivity is the hallmark of the particular ligation architecture, which is distinct from previous ligation architectures designed as “nick ligations”. Interestingly, the fidelity of the native chemical ligation of peptide nucleic acids was increased by more than one order of magnitude when performing the ligation in such a way that an abasic‐site mimic was formed opposite an unpaired template base. It is shown that the high sequence fidelity of the abasic ligation could facilitate the MALDI‐TOF mass‐spectrometric analysis of early cancer onset by allowing the detection of as little as 0.2 % of single‐base mutant DNA in the presence of 99.8 % wild‐type DNA.