Author:

Keywords:

Biopolymers, Cyclohexanes, DNA, DNA-Directed DNA Polymerase, Directed Molecular Evolution, Kinetics, Nucleotides, Templates, Genetic, 5'-triphosphates, substrate, oligonucleotides, Science & Technology, Life Sciences & Biomedicine, Biochemistry & Molecular Biology, REVERSE-TRANSCRIPTASE, FUNCTIONALIZED DNA, SUGAR RECOGNITION, GENETIC ALPHABET, POLYMERASE, RNA, NUCLEOSIDE, SUBSTRATE, 5'-TRIPHOSPHATES, TRIPHOSPHATE, 05 Environmental Sciences, 06 Biological Sciences, 08 Information and Computing Sciences, Developmental Biology, 31 Biological sciences, 34 Chemical sciences, 41 Environmental sciences

Abstract:

DNA polymerases from different evolutionary families [Vent (exo-) DNA polymerase from the B-family polymerases, Taq DNA polymerase from the A-family polymerases and HIV reverse transcriptase from the reverse transcriptase family] were examined for their ability to incorporate the sugar-modified cyclohexenyl nucleoside triphosphates. All enzymes were able to use the cyclohexenyl nucleotides as a substrate. Using Vent (exo-) DNA polymerase and HIV reverse transcriptase, we were even able to incorporate seven consecutive cyclohexenyl nucleotides. Using a cyclohexenyl nucleic acid (CeNA) template, all enzymes tested were also able to synthesize a short DNA fragment. Since the DNA-dependent CeNA polymerization and the CeNA-dependent DNA polymerization is possible to a limited extend, we suggest CeNA as an ideal candidate to use in directed evolution methods for the development of a polymerase capable of replicating CeNA.