Enzymatic synthesis of DNA strands containing α-L-LNA (α-L-configured locked nucleic acid) thymine nucleotides.
We describe the first enzymatic incorporation of an α-L-LNA nucleotide into an oligonucleotide. It was found that the 5'-triphosphate of α-L-LNA is a substrate for the DNA polymerases KOD, 9°N(m), Phusion and HIV RT. Three dispersed α-L-LNA thymine nucleotides can be incorporated into DNA strands by all four polymerases, but they were unable to perform consecutive incorporations of α-L-LNA nucleotides. In addition it was found that primer extension can be achieved using templates containing one α-L-LNA nucleotide.
Department of Physics, Chemistry and Pharmacy, Nucleic Acid Center, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
Torben
T
Højland
Rakesh N
RN
Veedu
Birte
B
Vester
Jesper
J
Wengel
D004247:DNA / Q000737:chemistry*; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry*; D009841:Oligonucleotides / Q000737:chemistry*; D013942:Thymine Nucleotides / Q000737:chemistry*
D009841:Oligonucleotides; D013942:Thymine Nucleotides; C477371:locked nucleic acid; D004247:DNA; D004259:DNA-Directed DNA Polymerase