Abstract:
:The nature of mammalian origins of DNA replication remains controversial and this is primarily because two-dimensional gel replicon mapping techniques have identified broad zones of replication initiation whereas several other techniques, such as quantitative PCR, have disclosed more discrete sites of initiation at the same chromosomal loci. In this report we analyze the replication of an amplified genomic region encompassing the 3'-end of the GNAI3 gene, the entire GNAT2 gene and the intergenic region between them in exponentially growing Chinese hamster fibroblasts. These cells express GNAI3 but not GNAT2 . The replication pattern was first analyzed by two-dimensional neutral-alkaline gel electrophoresis. Surprisingly, the results revealed a small preferential zone of replication initiation, of at most 1.7 kb, located in a limited part of the GNAI3 - GNAT2 intergenic region. Mapping of this initiation zone was then confirmed by quantitative PCR. The agreement between the two techniques exploited here strengthens the hypothesis that preferred sites of replication initiation do exist in mammalian genomes.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Toledo F,Baron B,Fernandez MA,Lachagès AM,Mayau V,Buttin G,Debatisse Mdoi
10.1093/nar/26.10.2313subject
Has Abstractpub_date
1998-05-15 00:00:00pages
2313-21issue
10eissn
0305-1048issn
1362-4962pii
gkb401journal_volume
26pub_type
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