Počet záznamů: 1
2‐Formyl‐dATP as Substrate for Polymerase Synthesis of Reactive DNA Bearing an Aldehyde Group in the Minor Groove
- 1.0525595 - ÚOCHB 2021 RIV DE eng J - Článek v odborném periodiku
Krömer, Matouš - Brunderová, Mária - Ivancová, Ivana - Poštová Slavětínská, Lenka - Hocek, Michal
2‐Formyl‐dATP as Substrate for Polymerase Synthesis of Reactive DNA Bearing an Aldehyde Group in the Minor Groove.
ChemPlusChem. Roč. 85, č. 6 (2020), s. 1164-1170. ISSN 2192-6506. E-ISSN 2192-6506
Grant CEP: GA ČR(CZ) GA18-03305S; GA MŠMT(CZ) EF16_019/0000729
Grant ostatní: AV ČR(CZ) AP1501
Program: Akademická prémie - Praemium Academiae
Institucionální podpora: RVO:61388963
Klíčová slova: bioconjugations * DNA polymerases * nucleotides * peptides * reductive amination
Obor OECD: Organic chemistry
Impakt faktor: 2.863, rok: 2020
Způsob publikování: Omezený přístup
https://chemistry-europe.onlinelibrary.wiley.com/doi/full/10.1002/cplu.202000287
2‐Formyl‐2′‐deoxyadenosine triphosphate (dCHOATP ) was synthesized and tested as a substrate in enzymatic synthesis of DNA modified in the minor groove with a reactive aldehyde group. The multistep synthesis of dCHOATP was based on the preparation of protected 2‐dihydroxyethyl‐2′‐deoxyadenosine intemediate, which was triphosphorylated and converted to aldehyde through oxidative cleavage. The dCHOATP triphosphate was a moderate substrate for KOD XL DNA polymerase, and was used for enzymatic synthesis of some sequences using primer extension (PEX). On the other hand, longer sequences (31‐mer) with higher number of modifications, or sequences with modifications at adjacent positions did not give full extension. Single‐nucleotide extension followed by PEX was used for site‐specific incorporation of one aldehyde‐linked adenosine into a longer 49‐mer sequence. The reactive formyl group was used for cross‐linking with peptides and proteins using reductive amination and for fluorescent labelling through oxime formation with an AlexaFluor647‐linked hydroxylamine.
Trvalý link: http://hdl.handle.net/11104/0309707
Počet záznamů: 1