Premium
Chemical Morphing of DNA Containing Four Noncanonical Bases
Author(s) -
Eremeeva Elena,
Abramov Michail,
Margamuljana Lia,
Rozenski Jef,
Pezo Valerie,
Marlière Philippe,
Herdewijn Piet
Publication year - 2016
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201601529
Subject(s) - polymerase , nucleotide , nucleobase , dna polymerase , dna , chemistry , dna replication , template , biochemistry , computational biology , combinatorial chemistry , gene , biology , nanotechnology , materials science
The ability of alternative nucleic acids, in which all four nucleobases are substituted, to replicate in vitro and to serve as genetic templates in vivo was evaluated. A nucleotide triphosphate set of 5‐chloro‐2′‐deoxyuridine, 7‐deaza‐2′‐deoxyadenosine, 5‐fluoro‐2′‐deoxycytidine, and 7‐deaza‐2′deoxyguanosine successfully underwent polymerase chain reaction (PCR) amplification using templates of different lengths (57 or 525mer) and Taq or Vent ( exo ‐) DNA polymerases as catalysts. Furthermore, a fully morphed gene encoding a dihydrofolate reductase was generated by PCR using these fully substituted nucleotides and was shown to transform and confer trimethoprim resistance to E. coli . These results demonstrated that fully modified templates were accurately read by the bacterial replication machinery and provide the first example of a long fully modified DNA molecule being functional in vivo.