Wu A F, Wu T T
Prep Biochem. 1982;12(1):1-10. doi: 10.1080/00327488208065546.
Commercially available calf thymus DNA could be fractionated into different populations by DEAE-cellulose column chromatography, presumably due to the tertiary folding of the solid fibrous form of DNA which opened up gradually in solution. Depending on the duration in solution or the concentration of the solution, various peaks could be obtained by eluting the DEAE column with 0.01 M sodium phosphate buffer at pH 7.0 and 0.01, 0.4, 0.5, 0.6 and 1.0 M NaCl in the same buffer. The DNA fraction in each peak gave very narrow bands on 1% agarose gel electrophoresis, probably reflecting the folding and thus shielding of charges rather than molecular weight. The present fractionation procedure might be useful in characterizing the complicated network structure of calf thymus DNA and in studying the unfolding of DNA tertiary structure.
市售的小牛胸腺DNA可通过DEAE - 纤维素柱色谱法分离成不同的组分,这可能是由于DNA固体纤维形式的三级折叠在溶液中逐渐展开所致。根据在溶液中的持续时间或溶液浓度,用pH 7.0的0.01 M磷酸钠缓冲液以及该缓冲液中0.01、0.4、0.5、0.6和1.0 M的NaCl洗脱DEAE柱,可获得不同的峰。每个峰中的DNA组分在1%琼脂糖凝胶电泳上呈现出非常窄的条带,这可能反映了折叠以及由此产生的电荷屏蔽,而非分子量。目前的分级分离方法可能有助于表征小牛胸腺DNA复杂的网络结构,并研究DNA三级结构的展开。