oriGNAI3: a narrow zone of preferential replication initiation in mammalian cells identified by 2D gel and competitive PCR replicon mapping techniques.

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 Res

journal_title

Nucleic acids research

authors

Toledo F,Baron B,Fernandez MA,Lachagès AM,Mayau V,Buttin G,Debatisse M

doi

10.1093/nar/26.10.2313

subject

Has Abstract

pub_date

1998-05-15 00:00:00

pages

2313-21

issue

10

eissn

0305-1048

issn

1362-4962

pii

gkb401

journal_volume

26

pub_type

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