A cell-free replication system and uses thereof

Details A cell-free replication system and uses thereof A cell-free replication system and uses thereof

C - Chemistry, Metallurgy

12

N

C12N 15/10 (2006.01) C07K 14/47 (2006.01) C12N 5/00 (2006.01) C12N 9/12 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2302480

The invention relates to improvements in cell-free systems for initiating DNA replication under cell cycle control. The system comprises a synchronous population of G1 nuclei which have been released from G0; and S phase cytosol. In a preferred aspect, a polypeptide is supplied to the system, wherein the polypeptide is Cdc6 and/or at least one MCM protein. The system is suitable for DNA replication assays, for example to test substances which modulate DNA replication.

Cette invention traite d'améliorations apportées à des systèmes acellulaires dans le but de déclencher une réplication d'ADN conformément au cycle cellulaire. Ce système possède une population synchrone de noyaux en phase G1 qui ont été libérés lors de la phase G0 et un cytosol en phase S. Selon un mode de réalisation préféré, un polypeptide est introduit dans ce système, ce polypeptide étant une protéine Cdc6 et/ou au moins une protéine MCM. Ce système convient à des analyses concernant la réplication d'ADN, par exemple, pour déceler des substances qui modulent la réplication d'ADN.

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