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PMID: 7935453 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mapping an initiation region of DNA replication at a single-copy chromosomal locus in Drosophila melanogaster cells by two-dimensional gel methods and PCR-mediated nascent-strand analysis: multiple replication origins in a broad zone.

Molecular and cellular biology ·Vol. 14 ·No. 11 ·1994-11-00 ·Pages 7394-403

Shinomiya T, Ina S

Abstract

We have mapped an initiation region of DNA replication at a single-copy chromosomal locus in exponentially proliferating Drosophila tissue culture cells, using two-dimensional (2D) gel replicon mapping methods and PCR-mediated analysis of nascent strands. The initiation region was first localized downstream of the DNA polymerase alpha gene by determining direction of replication forks with the neutral/alkaline 2D gel method. Distribution of replication origins in the initiation region was further analyzed by using two types of 2D gel methods (neutral/neutral and neutral/alkaline) and PCR-mediated nascent-strand analysis. Results obtained by three independent methods were essentially consistent with each other and indicated that multiple replication origins are distributed in a broad zone of approximately 10 kb. The nucleotide sequence of an approximately 20-kb region that encompasses the initiation region was determined and searched for sequence elements potentially related to function of replication origins.

MeSH Terms
Animals Base Sequence DNA/genetics DNA Replication/genetics Drosophila melanogaster/genetics Electrophoresis, Gel, Two-Dimensional Molecular Sequence Data Polymerase Chain Reaction Replication Origin/genetics Restriction Mapping
Chemicals
DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Shinomiya T
Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
Ina S
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.11.7394-7403.1994
Published
1994-11-00
Pages
7394-403
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC359274
Subset
IM
Databases
GENBANK
D28563
External Links
PubMed source DOI: 10.1128/mcb.14.11.7394-7403.1994 PMC Full Text View journal details
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