DNA Synthesis in Isolated HeLa Nuclei: A System for Continuation of Replication in vivo
1973
Hershey, Howard V. | Stieber, Joseph F. | Mueller, Gerald C.
A nuclear system from synchronized HeLa cells is described which continues DNA replication at sites which were active in vivo. Under the defined optimal conditions the initial rates of DNA replication in isolated nuclei range from 50 to 100% of that in the cells of origin; 1--2% of the HeLa DNA can be replicated in vitro. Proteins of the cytosol are essential to the stabilization of the replication system. Experiments in which the parental strands were fractionally labelled with bromodeoxyuridine provide evidence that the ordered nature of DNA replication in vivo is preserved in the system in vitro. Repair replication of homopolymer synthesis was insignificant under the conditions of study.
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