Purification of mouse H1 histones expressed in Escherichia coli.
We amplified the coding regions of the previously cloned H1 genes H1-1, H1 zero and H1t and inserted them into the expression vector pET-11d. The synthesis of the H1 histones can be induced in the appropriate strains of bacteria, and the H1 histones can be readily purified. We report detailed protocols for the purification of the expressed proteins using combinations of ion-exchange and reverse-phase HPLC. Sufficient amounts of each pure variant protein can be obtained for use in physical studies of H1-DNA interactions.
Wellman
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SE
Department of Pharmacology and Toxicology, University of Mississippi Medical Center, Jackson 39216-4505, USA.
Song
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Y
Su
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Mamoon
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NM
D000818:Animals; D001483:Base Sequence; D001709:Biotechnology; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D006657:Histones / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D066298:In Vitro Techniques; D051379:Mice; D011485:Protein Binding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
D017931:DNA Primers; D006657:Histones; D011994:Recombinant Proteins; D004247:DNA