Study of DNA replication in Drosophila using cell free in vitro system.
Study of DNA replication in Drosophila using cell free in vitro system.
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DOI：
10.4161/cc.9.4.10730
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发表时间：
2010-02-15
期刊：
影响因子：
--
通讯作者：
Chesnokov I
中科院分区：
文献类型：
--
作者：
Svitin A;Chesnokov I
Using Drosophila early egg extracts we have developed an optimized cell free system to study DNA replication. The efficiency of replication depends on a cold treatment of Drosophila embryos before the extract preparation and a formation of nuclei facilitated by the addition of membrane fractions to the extracts. In vitro DNA replication is ORC and CDC6 dependent, as a removal of these proteins from the extracts abolishes DNA replication. The N-terminal part of Orc1 protein, which is important for non-replicative functions of ORC, is dispensable for the replication in vitro. We also show that the conserved ATPase motif of CDC6 is crucial for the replication. Our studies indicate that a Drosophila cell free system proves to be an extremely useful tool for a functional dissection of the processes and factors involved in DNA replication in metazoans.
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