Publication details

DNA interaction with zinc(II) ions

Authors

LUKÁŠ Nejdl RUTTKAY-NEDECKÝ Branislav KUDR Jiří KŘÍŽKOVÁ Soňa ŠMERKOVÁ Kristýna DOSTÁLOVÁ Simona VACULOVIČOVÁ Markéta KOPEL Pavel ZEHNÁLEK Josef TRNKOVÁ Libuše BABULA Petr ADAM Vojtěch KIZEK René

Year of publication 2014
Type Article in Periodical
Magazine / Source International journal of biological macromolecules
MU Faculty or unit

Faculty of Science

Citation
Web http://www.sciencedirect.com/science/article/pii/S0141813013006636
Doi http://dx.doi.org/10.1016/j.ijbiomac.2013.12.013
Field Physical chemistry and theoretical chemistry
Keywords UV/VIS spectrophotometry; gel electrophoresis; melting temperature
Description We focused on interactions of Zn(II) with DNA in this study. These interactions were monitored using UV/vis spectrophotometry and gel electrophoresis. Firstly, we isolated and amplified 498 bp fragment of DNA. Samples were obtained by incubation of DNA fragment with Zn(II) for 60 min at 25 degrees C. After incubation, the samples were dialyzed and analyzed immediately. In this way, DNA was converted into a metal bound DNA (Zn-DNA). Interaction of Zn(II) with DNA caused change in the absorption spectrum (190-350 nm) and decrease in the melting temperature (Tm) of Zn-DNA. Spectrophotometric (UV/vis) analysis showed that increasing concentrations of zinc(II) ions led to the increase in the absorbance at 200 nm and decrease in absorbance at 251 nm. Application of zinc(II) ions at 5.5 mu M concentration caused decrease in Tm for app. 7.5 degrees C in average in comparison with control (75.5 +/- 3 degrees C). The lowest melting temperature (60.5 +/- 2.5 degrees C) was observed after application of zinc(II) ions at 33 mu M concentration. Gel electrophoresis proved significance of Zn(II) in the renaturation of DNA. Samples of Zn-DNA (15 mu M DNA + 5.5-55 mu M Zn(II)) caused significant changes in the renaturation of DNA in comparison with the control, untreated DNA (15 mu M DNA). (C) 2013 Elsevier B.V. All rights reserved.

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