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Base‐Modified Nucleic Acids as a Powerful Tool for Synthetic Biology and Biotechnology
Author(s) -
Eremeeva Elena,
Abramov Michail,
Margamuljana Lia,
Herdewijn Piet
Publication year - 2017
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201700679
Subject(s) - oligonucleotide , taq polymerase , polymerase , nucleic acid , restriction enzyme , dna , biology , biochemistry , endonuclease , nucleoside triphosphate , polymerase chain reaction , base pair , computational biology , nucleoside , dna polymerase , microbiology and biotechnology , nucleotide , chemistry , gene , thermus aquaticus
The ability of various nucleoside triphosphate analogues of deoxyguanosine and deoxycytidine with 7‐deazadeoxyadenosine (A 1 ) and 5‐chlorodeoxyuridine (T 1 ) to serve as substrates for Taq DNA polymerase was evaluated. The triphosphate set composed of A 1 , T 1 , and 7‐deazadeoxyguanosine with either 5‐methyldeoxycytidine or 5‐fluorodeoxycytidine was successfully employed in the polymerase chain reaction (PCR) of 1.5 kb fragments as well as random oligonucleotide libraries. Another effective combination of triphosphates for the synthesis of a 1 kb PCR product was A 1 , T 1 , deoxyinosine, and 5‐bromodeoxycytidine. In vivo experiments using an antibiotic‐resistant gene containing the latter set demonstrated that the bacterial machinery accepts fully modified sequences as genetic templates. Moreover, the ability of the base‐modified segments to selectively protect DNA from cleavage by restriction endonucleases was shown. This approach can be used to regulate the endonuclease cleavage pattern.