Simplified Method for Rapid Purification of Soluble Histones (CROSBI ID 238329)
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Podaci o odgovornosti
Ivić, Nives ; Groschup, Bernhard ; Bilokapić, Silvija ; Halić, Mario
engleski
Simplified Method for Rapid Purification of Soluble Histones
Functional and structural studies of histone-chaperone complexes, nucleosome modifications, their interactions with remodelers and regulatory proteins rely on obtaining recombinant histones from bacteria. In the present study, we show that co-expression of Xenopus laevis histone pairs leads to production of soluble H2AH2B heterodimer and (H3H4)2 heterotetramer. The soluble histone complexes are purified by simple chromatographic techniques. Obtained H2AH2B dimer and H3H4 tetramer are proficient in histone chaperone binding and histone octamer and nucleosome formation. Our optimized protocol enables rapid purification of multiple soluble histone variants with a remarkable high yield and simplifies histone octamer preparation. We expect that this simple approach will contribute to the histone chaperone and chromatin research.
histone ; nucleosome ; chromatin ; protein co-expression
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