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Author:
Date:
2014-03-20
[Abstract] Helicases are a class of enzymes which are motor proteins using energy derived from ATP hydrolysis to move directionally along a nucliec acid phosphodiester backbone (such as DNA, RNA and DNA-RNA hybrids) and separate two annealed nucleic acid strands. Many cellular processes, such as transcription, DNA replication, recombination and DNA repair involve helicase activity. Here, we provide a protocol to analyze helicase activities in vitro. In this protocol, the DNA helicase protein Merkel cell polyomavirus large T-antigen was expressed in the mammalian cell line HEK293 and immoblized on an IgG resin. The helicase assay is performing while the protein is immoblized on IgG resin.
[摘要] 解旋酶是一类酶，其是使用来自ATP水解的能量沿着核酸磷酸二酯主链（例如DNA，RNA和DNA-RNA杂交体）定向移动并分离两条退火的核酸链的运动蛋白。 许多细胞过程，例如转录，DNA复制，重组和DNA修复涉及解旋酶活性。 在这里，我们提供了一个协议，以分析解旋酶活性在体外。 在该协议中，DNA解旋酶蛋白Merkel细胞多瘤病毒大T抗原在哺乳动物细胞系HEK293中表达并固定在IgG树脂上。 进行解旋酶测定，同时蛋白质固定在IgG树脂上。
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