Polycistronic coexpression and nondenaturing purification of histone octamers

Histone octamers are the basic building blocks of chromatin and are platforms for diverse genetic mechanisms. We report a simple method for preparing recombinant histone octamers by overexpressing all four histones from a single polycistronic vector followed by standard chromatography under native c...

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Bibliographic Details
Published in:Analytical biochemistry 2012-08, Vol.427 (2), p.190-192
Main Authors: Shim, Yoonjung, Duan, Ming-Rui, Chen, Xuejing, Smerdon, Michael J., Min, Jung-Hyun
Format: Article
Language:English
Subjects:
Bacteria
Bacterial Proteins - genetics
Bacterial Proteins - isolation & purification
Bacterial Proteins - metabolism
Chromatin
Chromatin - genetics
Chromatin - metabolism
chromatography
Chromatography, Gel
Electrophoresis, Polyacrylamide Gel
Gene Expression
Genetic Vectors - chemistry
Genetic Vectors - genetics
Histone
histones
Histones - genetics
Histones - isolation & purification
Histones - metabolism
Nucleosome
Nucleosomes - genetics
Nucleosomes - metabolism
Polycistronic vector
Protein Isoforms - genetics
Protein Isoforms - isolation & purification
Protein Isoforms - metabolism
Protein Multimerization
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Recombinant Proteins - metabolism
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Summary:Histone octamers are the basic building blocks of chromatin and are platforms for diverse genetic mechanisms. We report a simple method for preparing recombinant histone octamers by overexpressing all four histones from a single polycistronic vector followed by standard chromatography under native conditions. This approach reduces the time needed for the octamer preparation to a single day and should be applicable to making a variety of unmodified and modified histone octamers.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2012.05.006