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酸提取对成熟禽红细胞DNA模板活性影响的原位研究。

In situ studies on the effect of acid extraction on the DNA template activity of mature avian erythrocytes.

作者信息

Klass M R

出版信息

Cell Differ. 1975 Mar;3(6):335-45. doi: 10.1016/0045-6039(75)90002-0.

Abstract

Exogenous E. coli RNA polymerase was used to determine the in situ DNA template activity of ethanol/acetone fixed avian erythrocytes. No RNA polymerase-catalyzed incorporation of 3H-UTP was detected in mature avian erythrocytes while simultaneously fixed avian lymphocytes did exhibit incorporation of 3H-UTP. Nuclei of mature erythrocytes which were subjected to treatments known to remove histones showed dramatic increases in RNA polymerase-catalyzed incorporation of 3H-UTP. The chromatin of treated cells was presumed to be more accessible to RNA polymerase as determined by the increase in RNA polymerase-catalyzed incorporation of 3H-UTP. Incubation of acid-treated nuclei in poly-L-lysine prior to incubation with RNA polymerase failed to inhibit the incorporation of 3H-UTP. Possible mechanisms for the inactivation of avian erythrocyte nuclei are discussed.

摘要

使用外源大肠杆菌RNA聚合酶来测定乙醇/丙酮固定的禽类红细胞的原位DNA模板活性。在成熟禽类红细胞中未检测到RNA聚合酶催化的3H-UTP掺入,而同时固定的禽类淋巴细胞确实表现出3H-UTP的掺入。经过已知可去除组蛋白的处理的成熟红细胞细胞核,其RNA聚合酶催化的3H-UTP掺入显著增加。根据RNA聚合酶催化的3H-UTP掺入增加判断,处理过的细胞的染色质被认为对RNA聚合酶更易接近。在用RNA聚合酶孵育之前,先将酸处理过的细胞核在聚-L-赖氨酸中孵育,未能抑制3H-UTP的掺入。讨论了禽类红细胞细胞核失活的可能机制。

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