Relationship between nicotinamide adenine dinucleotide concentration and in vitro synthesis of poly(adenosine diphosphate ribose) on purified nucleosomes.
Relationship between nicotinamide adenine dinucleotide concentration and in vitro synthesis of poly(adenosine diphosphate ribose) on purified nucleosomes.
复制标题
烟酰胺腺嘌呤二核苷酸浓度与纯化核小体上聚（腺苷二磷酸核糖）体外合成之间的关系。
DOI：
10.1021/bi00564a013
复制
发表时间：
1980
期刊：
影响因子：
2.9
通讯作者：
Smulson,M
中科院分区：
文献类型：
--
作者：
Butt,TR;Smulson,M
材料与方法材料：[32P] NAD要么从新英格兰核公司购买（比活度32-56 Ci/mmol），要么通过前面描述的程序合成（Jump等，1979）。e«me- 2,8 -3* H] NAD （3.4 Ci/mmol）也购自New England Nuclear。所有试剂均为分析级试剂。微球菌核酸酶购自Worthington公司，蛋白酶K购自merck公司。大肠杆菌核糖体亚基是来自多伦多大学扎希德博士的礼物。采用柯达SB5 x射线胶片进行自射线照相。在Spinner flask中37℃悬浮培养HeLa S3细胞。本研究中使用的所有细胞均采用异步培养至中等密度[（5-8）× 105个细胞/mL培养基]。HeLa核染色质的制备。采用Sporn等人（1969）的方法制备HeLa细胞核，用至少含缓冲液的Triton X-100（0.3%）洗涤
Materials and MethodsMaterials.[32P] NAD was either purchased from New England Nuclear (specific activity 32-56 Ci/mmol) or synthesized by the procedure described earlier (Jump et al., 1979).[at? e «me-2, 8-3* H] NAD (3.4 Ci/mmol) was also purchased from New England Nuclear. All the reagents were of ana-lytical grade. Micrococcal nuclease was purchased from Worthington and Proteinase K fromMerck. Escherichia coli ribosomal subunits were a gift from Dr. Zahid of the Univ-ersity of Toronto. Kodak SB5 X-ray film was used for au-toradiography. HeLa S3 cells were maintained in suspension cultures at 37 C in Spinner flasks. All the cells used in the present studies were grown asynchronously to a midlog density [(5-8) X 105 cells/mL of culture medium]. Preparation of Chromatin from HeLa Nuclei. HeLa cell nuclei were prepared by the method of Sporn et al.(1969) and washed with Triton X-100 (0.3%) containing buffer at least