A method of immobilizing genomic DNA on microcrystalline cellulose was described to isolate DNA-binding
proteins. At first, DNA fragments were prepared by sonication and immobilized on cellulose phase. After the
immobilization, DNA duplex formation was done. Using this microcrystalline cellulose affinity chromatography
technique, DNA-binding proteins from kumquat (Fortunella margarita Swingle) leaf samples were isolated and then
analyzed by Liquid Chromatography–Mass Spectrometry (LC-MS/MS). LC-MS/MS analysis showed that twenty-eight
kinds of protein mainly including histones, protein-synthetic proteins and other DNA-binding proteins were identified.
The identification list consists with the results in previous research on DNA-binding proteins isolation. It further suggests
that the technique developed in this study can be applied to the effective isolation of DNA-binding proteins.
Keywords: Affinity chromatography, DNA-binding proteins, DNA duplex formation, DNA immobilization, microcrystalline
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