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Vol 46(2012) N 4 p. 596-604; A.Y. Rubina*, M.V. Tsybulskaya, O.A. Zasedateleva, S.B. Popletaeva, M.A. Filippova, V.I. Butvilovskaya, S.A. Surzhikov, A.A. Stomakhin, A.S. Zasedatelev Sequence Specificity of Protein-Oligonucleotide Interactions and Protein Isolation on Sorptive Media Containing Immobilized Protein Specific Oligonucleotides Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia*arubina@biochip.ru Received - 2011-11-02; Accepted - 2011-12-22 The sequence specificity of protein-oligonucleotide interactions based on the example of binase interaction with oligodeoxyribonucleotides immobilized in biochip gel elements has been studied. Constants of the preferable binding of binase to the selected nucleotide sequences were measured. The GAGAGAG and GAGAGAGAG oligodeoxyribonucleotides, which specifically bind to binase, were used as molecular probes to develop affine sorptive media for binase isolation and concentration from diluted water solutions. The volume capacity of affinity sorbents with immobilized oligodeoxyribonucleotides was found to be 2.6 and 2.3 mg of binase per 1 mL of sorbent for GAGAGAG and GAGAGAGAG, respectively. It was shown that, after affinity chromatography and elution from sorptive media, the oligonucleotide specificity of binase corresponds to the specificity of the initial sample. biochip, hydrogel microarray, protein-oligodeoxyribonucleotide complexes, constants of binding, binase, oligodeoxyribonucleotides, affinity chromatography, protein concentrating, DNA-protein interactions |
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