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- Title
Chemical Morphing of DNA Containing Four Noncanonical Bases.
- Authors
Eremeeva, Elena; Abramov, Michail; Margamuljana, Lia; Rozenski, Jef; Pezo, Valerie; Marlière, Philippe; Herdewijn, Piet
- Abstract
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.
- Subjects
BASE pairs; NUCLEOSIDES; CHEMICAL research; NUCLEIC acids
- Publication
Angewandte Chemie, 2016, Vol 128, Issue 26, p7641
- ISSN
0044-8249
- Publication type
Article
- DOI
10.1002/ange.201601529