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碱性核酸内切酶在正常和受辐照小鼠胸腺、脾脏及肝脏细胞核中可溶性染色质释放过程中的作用

Role of alkaline endonucleases in the release of soluble chromatin from thymus, spleen and liver nuclei of normal and irradiated mice.

作者信息

Suciu D

出版信息

Int J Radiat Biol Relat Stud Phys Chem Med. 1979 Feb;35(2):119-31. doi: 10.1080/09553007914550141.

Abstract

Thymus, spleen and liver nuclei released a large fraction of soluble chromatin in vitro when incubation was carried out in sucrose media containing low concentrations of CaCl2 and/or MgCl2. A significant fraction of deoxyribopolynucleotides (DPN) was also extracted from nuclei. After 30 min of incubation at 37 degrees C, the maximum release of soluble chromatin was observed near a pH of 8, which corresponds to the optimum pH of the alkaline endonuclease activity from thymus, spleen and liver. The soluble chromatin and DPN were precipitated by increasing the bivalent ion concentration of the medium. The protein/DNA ratio and the molecular weight of DNA suggest that the soluble chromatin and DPN represent nucleosome-like particles. The release of soluble chromatin in the first 4 hours of incubation was significantly increased if the nuclear fraction was isolated from the thymus and spleen of whole-body irradiated mice (1000 rad). This effect was absent in the liver nuclei.

摘要

当在含有低浓度氯化钙和/或氯化镁的蔗糖培养基中进行孵育时,胸腺、脾脏和肝脏的细胞核在体外释放出大部分可溶性染色质。还从细胞核中提取了相当一部分脱氧核糖多聚核苷酸(DPN)。在37℃孵育30分钟后,在pH值接近8时观察到可溶性染色质的最大释放,这与胸腺、脾脏和肝脏中碱性核酸内切酶活性的最佳pH值相对应。通过增加培养基中二价离子浓度可使可溶性染色质和DPN沉淀。蛋白质/DNA比值和DNA分子量表明,可溶性染色质和DPN代表核小体样颗粒。如果从全身照射(1000拉德)小鼠的胸腺和脾脏中分离核部分,则孵育最初4小时内可溶性染色质的释放会显著增加。肝脏细胞核中不存在这种效应。

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