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[通过聚丙烯酰胺凝胶中的蛋白质复性方法研究脱氧核糖核酸酶在哺乳动物细胞中的表达]

[Study of expression of desoxyribonucleases in mammalian cells by a protein renaturation method in polyacrylamide gel].

作者信息

Kit Iu Ia, Shuvaieva H Iu, Drel' V R, Ihumentseva N I, Drobot L B

机构信息

Institute of Cell Biology, NAS of Ukraine, Lviv.

出版信息

Ukr Biokhim Zh (1999). 2002 May-Jun;74(3):116-9.

Abstract

Analysis of enzymatic activity in polyacrylamide gel is based on highly effective separation of proteins by SDS-electrophoresis with their subsequent renaturation and detection of enzymatic activity. This method was used to study an expression of DNAases in culturing of cells HEK293, NIH 3T3, U937. We have found that in HEK293 cells the nucleases with molecular weights 47 and 45 kDa were expressed. The localization of DNAases in the cell nuclei was shown as well. Induction of apoptosis in HEK293 cells increase the level of p47 DNAase and causes the expression of novel 50 kDa DNAase. We suggested that those discovered DNAases could take part in apoptotic DNA degradation.

摘要

聚丙烯酰胺凝胶中酶活性的分析基于通过SDS-电泳对蛋白质进行高效分离,随后使其复性并检测酶活性。该方法用于研究HEK293、NIH 3T3、U937细胞培养中DNA酶的表达。我们发现,在HEK293细胞中表达了分子量为47 kDa和45 kDa的核酸酶。还显示了DNA酶在细胞核中的定位。HEK293细胞中凋亡的诱导增加了p47 DNA酶的水平,并导致了新的50 kDa DNA酶的表达。我们认为,那些发现的DNA酶可能参与凋亡性DNA降解。

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