Rapid detection and purification of sequence specific DNA binding proteins using magnetic separation | Zendy

Marija Mojsin | Zendy; Jelena Djurovic | Zendy; Isidora Petrović | Zendy; Aleksandar Krstić | Zendy; Danijela Drakulić | Zendy; Tijana Savić | Zendy; Milena Stevanović | Zendy

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Rapid detection and purification of sequence specific DNA binding proteins using magnetic separation

Author(s) -

Marija Mojsin,

Jelena Djurovic,

Isidora Petrović,

Aleksandar Krstić,

Danijela Drakulić,

Tijana Savić,

Milena Stevanović

Publication year - 2006

Publication title -

journal of the serbian chemical society

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.227

H-Index - 45

eISSN - 1820-7421

pISSN - 0352-5139

DOI - 10.2298/jsc0602135m

Subject(s) - biotinylation , recombinant dna , dna , streptavidin , chemistry , flag tag , microbiology and biotechnology , tandem affinity purification , biochemistry , protein purification , target protein , dna sequencing , chromatography , affinity chromatography , biology , biotin , fusion protein , gene , enzyme

In this paper, a method for the rapid identification and purification of se- quence specific DNA binding proteins based on magnetic separation is presented. This method was applied to confirm the binding of the human recombinant USF1 protein to its putative binding site (E-box) within the human SOX3 protomer. It has been shown that biotinylated DNA attached to streptavidin magnetic particles spe- cifically binds the USF1 protein in the presence of competitor DNA. It has also been demonstrated that the protein could be successfully eluted from the beads, in high yield and with restored DNA binding activity. The advantage of these procedures is that they could be applied for the identification and purification of any high-affinity sequence-specific DNA binding protein with only minor modifications.

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