Enriched autonomously replicating sequences in a nuclear matrix-DNA complex isolated from synchronized HeLa cells.
Nucleoids isolated from either synchronized or exponentially growing HeLa cells were digested with restriction enzymes to separate a nuclear matrix-bound DNA component from the rest. Partial libraries were constructed by inserting DNA fragments from both components into a yeast-bacteria plasmid vector. A random sample from these libraries was tested for ARS activity by a standard yeast transformation assay. We found that synchronization for DNA replication results in an enrichment for autonomously replicating sequences in the library constructed with the DNA component bound to the nuclear matrix.
D001483:Base Sequence; D002453:Cell Cycle; D002467:Cell Nucleus / Q000378:metabolism*; D004261:DNA Replication; D004262:DNA Restriction Enzymes; D004273:DNA, Neoplasm / Q000235:genetics*; D004274:DNA, Recombinant / Q000378:metabolism; D006367:HeLa Cells / Q000166:cytology; D006801:Humans; D007700:Kinetics; D010957:Plasmids
D004273:DNA, Neoplasm; D004274:DNA, Recombinant; D004262:DNA Restriction Enzymes