大鼠肝细胞核纯化、染色质分部及染色质非组蛋白双向凝胶电泳分析

Purification of Nuclei and Fractionation of Chromatin From rat Liver and Electrophoresis of Non-Histone Chromatin Proteins by two Dimensional Gel Electrophoresis

摘要: 本文使用改良的超离心法制备了大鼠肝细胞核，通过相差显微镜、电镜、转录活力及三种酶活力的测定，证实了所得细胞核较纯，其产率约为5%。使用羟基磷灰石（HAP）柱层析对染色质分部，并测定了各分部的紫外光谱。同时测定了核、染色质、染色质疏松结合非组蛋白（UP）、去up—染色质（UC）和HAP柱各分部的化学组成。对up和酚抽提法制得染色质紧密结合非组蛋白（NP）分别进行了双向凝胶电泳图谱分析。

Abstract: Nuclei were prepared from adult rat liver by a slightly modified ultracentrifugation method,with a yield of about 5%.The quality of rat liver nuclei has been ascertained by observations with the phase contract microscope,electron microscope,transcriptional activity and estimations of enzyme activity.The chromatin of rat liver was fractionated by the method of hydroxyapatite (HAP) chromatography.The U.V.spectrum of each fraction was obtained individually.The chemical compositions of the nuclei,chromatin,Up (a group of nonhistone proteins binding DNA loosly),deUpgchromatin and each fraction from HAP were determined.NP (a group of nonhistone proteins binding DNA tightly) were extracted from deUP-chromatin with phenol.UP and NP were analysed by two dimensional gel electrophoresis.