Enzymatic synthesis of DNA strands containing α-L-LNA (α-L-configured locked nucleic acid) thymine nucleotides

Torben Højland; Rakesh N Veedu; Birte Vester; Jesper Wengel

doi:10.4161/adna.19272

Enzymatic synthesis of DNA strands containing α-L-LNA (α-L-configured locked nucleic acid) thymine nucleotides

Artif DNA PNA XNA. 2012 Jan-Mar;3(1):14-21. doi: 10.4161/adna.19272.

Authors

Torben Højland¹, Rakesh N Veedu, Birte Vester, Jesper Wengel

Affiliation

¹ Department of Physics, Chemistry and Pharmacy, Nucleic Acid Center, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.

Abstract

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.

Keywords: enzymatic synthesis; modified nucleotide; polymerase; triphosphate; α-L-LNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA / chemistry*
DNA-Directed DNA Polymerase / chemistry*
Oligonucleotides / chemistry*
Thymine Nucleotides / chemistry*

Substances

Oligonucleotides
Thymine Nucleotides
locked nucleic acid
DNA
DNA-Directed DNA Polymerase