Abstract
The nuclear matrix is believed to contain sites of assembly of protein complexes that catalyze the initiation of DNA replication as well as DNA elongation. To explore this relationship, DNA replicated by human fibroblasts at the beginning of the S phase was purified and used to construct a cosmid library. Hybridization studies with a subgroup of clones (about one-sixth of the total clones in this library) showed that many of them were highlighted by probes prepared from early replicating DNA, as well as from nuclear matrix-associated DNA. Statistical analysis showed a positive correlation between these hybridization results. We seek to identify origins of replication that are activated early in the S phase of the cell cycle in human cells. Therefore, clones isolated from this library are being analyzed for the presence of structural motifs that have been found in other origins of replication and for potential sites of attachment to the nuclear matrix. This method of analysis is illustrated here using the published sequences for the origins of replication reported for the human lamin B2 and HPRT genes.
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