Open AccessDNA Synthesis in Isolated HeLa Nuclei
Author(s) -
Hershey Howard V.,
Stieber Joseph F.,
Mueller Gerald C.
Publication year - 1973
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1973.tb02770.x
Subject(s) - dna replication , hela , dna , control of chromosome duplication , replication (statistics) , bromodeoxyuridine , replication factor c , biology , eukaryotic dna replication , dna synthesis , in vitro , microbiology and biotechnology , replication protein a , semiconservative replication , genetics , dna binding protein , gene , virology , cell growth , transcription factor
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.