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PMID: 8041756 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Fine mapping of a replication origin of human DNA.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 91 ·No. 15 ·1994-07-19 ·Pages 7119-23

Giacca M, Zentilin L, Norio P, Diviacco S, Dimitrova D, Contreas G, Biamonti G, Perini G, Weighardt F, Riva S

Abstract

A highly sensitive procedure was developed for the identification of the origin of bidirectional DNA synthesis in single-copy replicons of mammalian cells. The method, which does not require cell synchronization or permeabilization, entails the absolute quantification, by a competitive PCR procedure in newly synthesized DNA samples, of the abundance of neighboring DNA fragments distributed along a given genomic region. This procedure was utilized for mapping the start site of DNA replication in a 13.7-kb region of human chromosome 19 coding for lamin B2, which is replicated immediately after the onset of S phase in HL-60 cells. Within this region, DNA replication initiates in a 474-bp area corresponding to the 3' noncoding end of the lamin B2 gene and the nontranscribed spacer between this gene and the 5' end of another highly transcribed one. This localization was obtained both in aphidicolin-synchronized and in exponentially growing HL-60 cells.

MeSH Terms
Animals Base Sequence Cell Line Cloning, Molecular DNA DNA Replication Humans Lamin Type B Lamins Molecular Sequence Data Nuclear Proteins/genetics Polymerase Chain Reaction/methods Simian virus 40/genetics
Chemicals
Lamin Type B Lamins Nuclear Proteins lamin B2 DNA
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Giacca M
International Centre for Genetic Engineering and Biotechnology, AREA Science Park, Trieste, Italy.
Zentilin L
Norio P
Diviacco S
Dimitrova D
Contreas G
Biamonti G
Perini G
Weighardt F
Riva S
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.91.15.7119
Published
1994-07-19
Pages
7119-23
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC44350
Subset
IM
Databases
GENBANK
M94363
External Links
PubMed source DOI: 10.1073/pnas.91.15.7119 PMC Full Text View journal details
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