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Cleavage of double stranded plasmid DNA by lanthanide complexes
Authors:Rittich Bohuslav  Spanová Alena  Falk Martin  Benes Milan J  Hrubý Martin
Affiliation:Department of Microbiology, Faculty of Science, Masaryk University Brno, Tvrdého 14, CZ-602 00 Brno, Czech Republic. rittich@sci.muni.cz
Abstract:The use of free lanthanide ions and their complexes for plasmid DNA pBR322 and chromosomal DNA cleavage was studied. Plasmid pBR322 DNA was treated by lanthanide chlorides (Eu(3+), La(3+), Nd(3+), Pr(3+), Gd(3+)) in HEPES buffer (pH 7.0, 7.5 and 8.0) at 24, 37, 50, 63, and 76 degrees C. The formation of linear and nicked plasmid forms was investigated depending on the reaction conditions. Heterogeneous lanthanide complexes of ethylenediamine tetraacetic acid (EDTA) immobilized on insoluble methacrylate support and iminodiacetic acid (IDA) immobilized on styrene support were used as catalysts plasmid for DNA pBR322 cleavage, too. The temperature of reaction mixture had substantial influence on cleavage rate. The precipitation of DNA occurred during the measurement of interactions between chromosomal DNA and La(3+) ions.
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