Study of DNA replication in Drosophila using cell free in vitro system

Academic Article

Abstract

  • 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. © 2010 Landes Bioscience.
  • Published In

  • Cell Cycle  Journal
  • Digital Object Identifier (doi)

    Pubmed Id

  • 3044113
  • Author List

  • Svitin A; Chesnokov I
  • Start Page

  • 815
  • End Page

  • 819
  • Volume

  • 9
  • Issue

  • 4