PROPERTIES OF FORMALDEHYDE-TREATED NUCLEOHISTONE
PROPERTIES OF FORMALDEHYDE-TREATED NUCLEOHISTONE
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DOI：
10.1021/bi00836a013
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发表时间：
1969-01-01
期刊：
影响因子：
2.9
通讯作者：
BONNER, J
中科院分区：
文献类型：
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作者：
BRUTLAG, D;SCHLEHUB.C;BONNER, J
材料与方法核组蛋白的制备。根据Bonner等人的方法从豌豆芽中制备染色质。（1968），不同之处在于初始研磨介质含有0.25 m蔗糖、0.01 m Tris（pH 8.0）和0.002 m MgCl 2。通过在Potter-Elvehjem均化器中的离心，将来自蔗糖纯化步骤的染色质沉淀重悬于40 ml 0.01 m三乙醇胺（pH 7.8; CalBiochem，不含重金属）中。将所得染色质溶液对0.01M三乙醇胺（pH 7.8）透析4小时。然后将染色质在Virtis匀浆器中在40 V下剪切90秒，在10，000 g下离心15分钟，并将上清液相对于0.01 m三乙醇胺（pH 7.8）透析过夜。所得核组蛋白通常具有20-40 OD 260的浓度，OD 230/OD 260的比率为约0.7，并且未显示出转化。
Materials and MethodsPreparation of Nucleohistone. Chromatin was prepared from pea budsaccording to the method of Bonner et al.(1968) except that the initial grinding media contained 0.25 m su-crose, 0.01 m Tris (pH 8.0), and 0.002 m MgCl2. The chromatin pellets from the sucrose purification step were resuspended in 40 ml of 0.01 m triethanolamine (pH 7.8; CalBiochem, heavy metal free) by homogenizing in a Potter-Elvehjem homogenizer. The resultingchromatin solution was dialyzed for 4 hr vs. 0.01 m triethanolamine (pH 7.8). The chromatin was then sheared in the Virtis homogenizer for 90 sec at 40 V, centrifuged at 10,000 g for 15 min, and the supernatant was dialyzed overnight vs. 0.01 m triethanolamine (pH 7.8). The resulting nucleohistone usually had a concentration of 20-40 OD26o, a ratio OD230/OD26o of about 0.7, and showed no tur-