Počet záznamů: 1
Anchored linear oligonucleotides: the effective tool for the real-time measurement of uracil DNA glycosylase activity
- 1.0547495 - ÚOCHB 2022 RIV GB eng J - Článek v odborném periodiku
Ligasová, A. - Rosenberg, Ivan - Bocková, Markéta - Homola, Jiří - Koberna, K.
Anchored linear oligonucleotides: the effective tool for the real-time measurement of uracil DNA glycosylase activity.
Open Biology. Roč. 11, č. 10 (2021), č. článku 210136. E-ISSN 2046-2441
Grant CEP: GA TA ČR(CZ) TN01000013; GA MŠMT(CZ) LM2018133; GA MZd NV15-31604A; GA ČR(CZ) GA19-02739S
Institucionální podpora: RVO:61388963 ; RVO:67985882
Klíčová slova: uracil DNA glycosylase * base excision repair * immobilized oligonucleotides * immobilized oligonucleotides * Förster resonance energy transfer * surface plasmon resonance
Obor OECD: Biochemistry and molecular biology; Optics (including laser optics and quantum optics) (URE-Y)
Impakt faktor: 7.000, rok: 2021
Způsob publikování: Open access
https://doi.org/10.1098/rsob.210136
Base excision repair is one of the important DNA repair mechanisms in cells. The fundamental role in this complex process is played by DNA glycosylases. Here, we present a novel approach for the real-time measurement of uracil DNA glycosylase activity, which employs selected oligonucleotides immobilized on the surface of magnetic nanoparticles and Förster resonance energy transfer. We also show that the approach can be performed by surface plasmon resonance sensor technology. We demonstrate that the immobilization of oligonucleotides provides much more reliable data than the free oligonucleotides including molecular beacons. Moreover, our results show that the method provides the possibility to address the relationship between the efficiency of uracil DNA glycosylase activity and the arrangement of the used oligonucleotide probes. For instance, the introduction of the nick into oligonucleotide containing the target base (uracil) resulted in the substantial decrease of uracil DNA glycosylase activity of both the bacterial glycosylase and glycosylases naturally present in nuclear lysates.
Trvalý link: http://hdl.handle.net/11104/0323709
Počet záznamů: 1