Expression of Rat Histone H1d inEscherichia coliand Its Purification

Histone H1 is involved in the folding of linear polynucleosomal filament into a 30-nm fiber. In an effort to understand the role of different domains of histone H1 in chromatin folding, we have now expressed rat histone H1d inEscherichia coliusing pTrc99A expression vector by providing a 6-His tag a...

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Published inProtein expression and purification Vol. 12; no. 1; pp. 38 - 44
Main Authors Bharath, M.M.Srinivas, Khadake, J.R., Rao, M.R.S.
Format Journal Article
LanguageEnglish
Published Elsevier Inc 01.02.1998
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Summary:Histone H1 is involved in the folding of linear polynucleosomal filament into a 30-nm fiber. In an effort to understand the role of different domains of histone H1 in chromatin folding, we have now expressed rat histone H1d inEscherichia coliusing pTrc99A expression vector by providing a 6-His tag at the C-terminus to facilitate its purification. The expressed protein histone H1d was purified from the soluble extract ofE. coliby employing Ni2+NTA–agarose and heparin–agarose chromatography. The recombinant histone H1d was shown to be authentic by its N-terminal amino acid analysis, its secondary structural characteristics, and its ability to (a) condense DNA and (b) bind specifically to synthetic four-way junction DNA.
ISSN:1046-5928
1096-0279
DOI:10.1006/prep.1997.0804