title: Synthesis of threose nucleic acid (TNA) triphosphates and oligonucleotides by polymerase-mediated primer extension.
abstract: This unit describes the chemical synthesis of α-L-threofuranosyl nucleic acid (TNA) triphosphates for thymidine (T), guanosine (G), cytidine (C), and the diaminopurine (D) analog of adenosine and their incorporation into TNA oligonucleotides by enzyme-mediated polymerization of a DNA primer-template complex. Starting from suitably protected threofuranosyl nucleosides, TNA triphosphates are synthesized in a single-pot reaction and purified by ion-exchange and HPLC chromatography. Purified TNA triphosphates are diluted into stock solutions and used as substrates for the synthesis of TNA oligonucleotides. Oligonucleotide synthesis is accomplished using Therminator DNA polymerase, a commercial variant of the 9(o)N DNA polymerase bearing the A485L mutation.
authors.affiliation: Center for Evolutionary Medicine and Informatics, The Biodesign Institute, and Department of Chemistry and Biochemistry at Arizona State University, Tempe, Arizona.
authors.forename: Su
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Center for Evolutionary Medicine and Informatics, The Biodesign Institute, and Department of Chemistry and Biochemistry at Arizona State University, Tempe, Arizona.
authors.forename: Hanyang
authors.initials: H
authors.lastname: Yu
authors.affiliation: Center for Evolutionary Medicine and Informatics, The Biodesign Institute, and Department of Chemistry and Biochemistry at Arizona State University, Tempe, Arizona.
authors.forename: John C
authors.initials: JC
authors.lastname: Chaput
mesh_terms: D003562:Cytidine / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry; D006151:Guanosine / Q000737:chemistry; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D011122:Polyphosphates / Q000737:chemistry*; D013780:Tetroses / Q000737:chemistry*; D013936:Thymidine / Q000737:chemistry
chemical_list: D017931:DNA Primers; D009841:Oligonucleotides; D011122:Polyphosphates; D013780:Tetroses; D006151:Guanosine; D003562:Cytidine; D004259:DNA-Directed DNA Polymerase; C005692:triphosphoric acid; D013936:Thymidine; C073321:erythrose