TY - JOUR
T1 - Additive-Free Enzymatic Phosphorylation and Ligation of Artificial Oligonucleotides with C-Nucleosides at the Reaction Points
AU - Oda, Yutaro
AU - Chiba, Junya
AU - Kurosaki, Fumihiro
AU - Yamade, Yusuke
AU - Inouye, Masahiko
N1 - Publisher Copyright:
© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
PY - 2019/8
Y1 - 2019/8
N2 - We report enzymatic phosphorylation and additive-free ligation of DNAs containing unnatural C-nucleotide residues through the action of T4 polynucleotide kinase and T4 DNA ligase. The artificial units are each made up of an alkynyl deoxyribose component and one of the unnatural nucleobases D*, T*, G*, and C*, corresponding—from a viewpoint of hydrogen-bonding patterns—to natural A, T, G, and C, respectively. Phosphorylation progressed quantitatively at the 5′-end in the cases of all of the artificial units in the chimeric DNAs. Ligation also smoothly progressed at the 5′-end in the cases of the D* and G* nucleotide residues, but only negligibly in those of their T* and C* counterparts. Chemical redesign of the last two units successfully improved the ligation efficiency, so that enzymatic ligation worked well for all of the artificial units in every 3′-natural⋅5′-artificial, 3′-artificial⋅5′-natural, and 3′-artificial⋅5′-artificial terminal combination at the nicks.
AB - We report enzymatic phosphorylation and additive-free ligation of DNAs containing unnatural C-nucleotide residues through the action of T4 polynucleotide kinase and T4 DNA ligase. The artificial units are each made up of an alkynyl deoxyribose component and one of the unnatural nucleobases D*, T*, G*, and C*, corresponding—from a viewpoint of hydrogen-bonding patterns—to natural A, T, G, and C, respectively. Phosphorylation progressed quantitatively at the 5′-end in the cases of all of the artificial units in the chimeric DNAs. Ligation also smoothly progressed at the 5′-end in the cases of the D* and G* nucleotide residues, but only negligibly in those of their T* and C* counterparts. Chemical redesign of the last two units successfully improved the ligation efficiency, so that enzymatic ligation worked well for all of the artificial units in every 3′-natural⋅5′-artificial, 3′-artificial⋅5′-natural, and 3′-artificial⋅5′-artificial terminal combination at the nicks.
KW - DNA
KW - DNA kinases
KW - alkynyl C-nucleosides
KW - ligases
KW - ligation
KW - phosphorylation
UR - http://www.scopus.com/inward/record.url?scp=85068234508&partnerID=8YFLogxK
U2 - 10.1002/cbic.201900217
DO - 10.1002/cbic.201900217
M3 - 学術論文
C2 - 30941833
AN - SCOPUS:85068234508
SN - 1439-4227
VL - 20
SP - 1945
EP - 1952
JO - ChemBioChem
JF - ChemBioChem
IS - 15
ER -