Fenske H, Eichhorn I, Böttger M, Lindigkeit R
Nucleic Acids Res. 1975 Oct;2(10):1975-85. doi: 10.1093/nar/2.10.1975.
It is shown that the release of the slightly lysine-rich histones f2a2 and f2b by 0.4 M ammonium sulfate from conventionally isolated chromatin is diminished in comparison to the lysed nuclei. The change in extractability is further demonstrated by the application of ethidium bromide. At a molar input ratio of 0.09 (moles ethidium bromide/moles nucleotide) and 0.4 M ammonium sulfate the slightly lysine-rich histones are released from the chromatin to 70 - 80% if the lysed nuclei are used. At 0.1 M ammonium sulfate ethidium bromide effected also a release of 50 % of histone f1. Comparable effects could not be observed with chromatin prepared in a conventional way but instead a tendency towards loss of histone f3 in the presence of ethidium bromide was observed.
结果表明,与裂解的细胞核相比,用0.4M硫酸铵从常规分离的染色质中释放出的富含赖氨酸的组蛋白f2a2和f2b减少。溴化乙锭的应用进一步证明了可提取性的变化。当溴化乙锭与核苷酸的摩尔输入比为0.09(溴化乙锭摩尔数/核苷酸摩尔数)且硫酸铵浓度为0.4M时,若使用裂解的细胞核,富含赖氨酸的组蛋白可从染色质中释放70%-80%。在硫酸铵浓度为0.1M时,溴化乙锭也能使50%的组蛋白f1释放。对于常规制备的染色质,未观察到类似效果,反而在溴化乙锭存在下观察到组蛋白f3有丢失的趋势。