Schröter H, Maier G, Ponstingl H, Nordheim A
EMBO J. 1985 Dec 30;4(13B):3867-72. doi: 10.1002/j.1460-2075.1985.tb04159.x.
Chicken erythrocyte nuclei were incubated with DNA intercalating agents in order to isolate from chromatin specific DNA-binding proteins whose binding specificity may be determined by DNA secondary and/or tertiary structure. The intercalating agents ethidium bromide (EtBr) and propidium iodide induce the specific release of high mobility group proteins HMG 14 and HMG 17 under low ionic strength conditions. Chloroquine (CQ) intercalation also results in the selective liberation of HMG 14 and HMG 17, but, in addition, selectively releases other nuclear proteins (including histone H1A) in a pH- and ionic strength-dependent fashion. The use of this new 'elutive intercalation' technique for the isolation and purification of 'sequence-specific' and 'helix-specific' DNA-binding proteins is suggested.
鸡红细胞核与DNA嵌入剂一起温育,以便从染色质中分离出特异性DNA结合蛋白,其结合特异性可能由DNA二级和/或三级结构决定。嵌入剂溴化乙锭(EtBr)和碘化丙啶在低离子强度条件下诱导高迁移率族蛋白HMG 14和HMG 17的特异性释放。氯喹(CQ)嵌入也导致HMG 14和HMG 17的选择性释放,但此外,还以pH和离子强度依赖性方式选择性释放其他核蛋白(包括组蛋白H1A)。建议使用这种新的“洗脱嵌入”技术来分离和纯化“序列特异性”和“螺旋特异性”DNA结合蛋白。