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成纤维细胞和淋巴细胞粗提物中的DNA连接酶活性。

DNA ligase activity in crude extracts of fibroblasts and lymphocytes.

作者信息

Saucier J M, Laval F

出版信息

Biochem Biophys Res Commun. 1983 Oct 31;116(2):657-62. doi: 10.1016/0006-291x(83)90575-2.

Abstract

DNA ligase activity was determined in crude cell extracts using a new assay which measures the retention of double stranded circular phage lambda DNA on nitrocellulose filters, and allows accurate determinations of the enzyme activity with cell concentration corresponding to 0.1 microgram of proteins. Using this assay, we show that the DNA ligase activity varies greatly among mammalian cell lines. The higher activity is found in actively growing fibroblasts where it is stimulated by dimethyl sulfate pretreatment of the cells, whereas the low activity measured in resting lymphocytes is not modified by dimethyl sulfate. The DNA ligase activity correlates with the cells sensitivity towards ionizing radiations.

摘要

使用一种新的检测方法测定粗细胞提取物中的DNA连接酶活性,该方法可测量双链环状噬菌体λDNA在硝酸纤维素滤膜上的保留情况,并能准确测定与0.1微克蛋白质相对应的细胞浓度下的酶活性。利用该检测方法,我们发现DNA连接酶活性在哺乳动物细胞系中差异很大。在活跃生长的成纤维细胞中活性较高,细胞经硫酸二甲酯预处理后活性会被刺激,而在静止淋巴细胞中测得的低活性不受硫酸二甲酯的影响。DNA连接酶活性与细胞对电离辐射的敏感性相关。

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