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Application of capillary electrophoresis for DNA-protein binding tests

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Abstract

The combination of affinity chromatography and capillary electrophoresis (CE-SDS) has been found to be a useful tool to analyse populations of proteins which specifically bind to ssor dsDNA. Proteins were extracted from tissue, cytosol or nuclei of meristems of Pisum sativum seedlings and separated on cellulose column functionalized with ss-, dsDNA (calf thymus) and ssDNA (P. sativum) at 2M concentration of sodium chloride. Electropherograms of the crude protein extracts show two fractions of proteins specific for dsDNA (calf thymus) and three fractions specific for ssDNA (calf thymus). Four and five fractions of proteins specific for ssDNA (P. sativum) were identified in the material isolated from cytosolic and nuclear extracts, respectively. Both ds- and ssDNA (calf thymus) form complexes with ca. 4.0 % of the total amount of proteins, while ssDNA (P. sativum) binds to ca. 11.0 % of cytosolic and 5.0 % of nuclear proteins.

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Abbreviations

CE:

capillary electrophoresis

DSB:

double-stranded

DNA:

binding protein

ds:

double-stranded

ss:

single-stranded

SSB:

single-stranded

DNA:

binding protein

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Authors and Affiliations

  1. Department of Cytophysiology, University of Łódź, Pilarskiego 14, PL-90231, Łódź, Poland

    Andrzej Kaźmierczak

  2. Department of Cytogenetics and Plant Molecular Biology, University of Łódź, Banacha 12/16, PL-90237, Poland

    Joanna M. Kaźmierczak

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  1. Andrzej Kaźmierczak
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  2. Joanna M. Kaźmierczak
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Kaźmierczak, A., Kaźmierczak, J.M. Application of capillary electrophoresis for DNA-protein binding tests. Acta Physiol Plant 25, 13–17 (2003). https://doi.org/10.1007/s11738-003-0031-2

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  • DOI: https://doi.org/10.1007/s11738-003-0031-2

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