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A Large Scale Procedure for Isolation of the Glycine-rich, Arginine-rich Histone and the Arginine-rich, Lysine-rich Histone in a Highly Purified Form

作者:Wesley C. Starbuck, Charles M. Mauritzen, Charles W. Taylor, Iengar S. Saroja, H. Busch · 发表于:Journal of Biological Chemistry · 年份:1968 · DOI:10.1016/s0021-9258(18)93546-5 · 被引用次数:52 · 研究领域:Enzyme Structure and Function、Genomics and Chromatin Dynamics、DNA and Nucleic Acid Chemistry

1. A procedure is presented for large scale fractionation of arginine-rich histones by exclusion chromatography. With this method, 2 to 3.5 g of histone Fractions 2a and 3 were fractionated. The two fractions contained the same histone components, but they differed in the amounts of each component. 2. In histone Fraction 2a, the glycine-rich, arginine-rich (GAR) histone accounted for approximately 70% of the protein present. This histone was obtained in a highly purified form after exclusion chromatography was repeated once, as shown by polyacrylamide gel electrophoresis, amino acid analysis, and peptide maps. Rechromatography of this product on Sephadex G-25 provided a GAR histone which was 96% pure. A quantity of GAR histone of 1 g was readily isolated by this procedure. 3. In histone Fraction 3, the arginine-rich, lysine-rich (AL) histone accounted for approximately 60% of the protein present. Rechromatography of the AL histone on Sephadex G-100 followed by rechromatography on Sephadex G-25 and Sephadex G-200 yielded a product approximately 96% pure. As in the case of the GAR histone, 1 g of the AL histone was readily obtained.