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PMID: 2542763 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Isolation of human sequences that replicate autonomously in human cells.

Molecular and cellular biology ·Vol. 9 ·No. 3 ·1989-03-00 ·Pages 1026-33

Krysan PJ, Haase SB, Calos MP

Abstract

We have isolated a heterogeneous collection of human genomic sequences which replicate autonomously when introduced into human cells. The novel strategy for the isolation of these sequences involved cloning random human DNA fragments into a defective Epstein-Barr virus vector. This vector alone was unable to replicate in human cells, but appeared to provide for the nuclear retention of linked DNA. The human sequences persist in a long-term replication assay (greater than 2 months) in the presence of the viral nuclear retention sequences. Using a short-term (4-day) assay, we showed that the human sequences are able to replicate in the absence of all viral sequences. The plasmids bearing human sequences were shown to replicate based on the persistence of MboI-sensitive plasmid DNA in the long-term assay and the appearance of DpnI-resistant DNA in the short-term assay. The human sequences were shown to be responsible for the replication activity and may represent authentic human origins of replication.

MeSH Terms
Base Sequence Cell Nucleus/metabolism Cloning, Molecular DNA Replication Genes Genetic Markers Genetic Vectors Herpesvirus 4, Human/genetics Humans Plasmids Replicon
Chemicals
Genetic Markers
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Krysan P J
Department of Genetics, Stanford University School of Medicine, California 94305.
Haase S B
Calos M P
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.9.3.1026-1033.1989
Published
1989-03-00
Pages
1026-33
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362692
Subset
IM
Grants
NCI NIH HHS · CA 33056 · United States
NCI NIH HHS · CA 44645 · United States
External Links
PubMed source DOI: 10.1128/mcb.9.3.1026-1033.1989 PMC Full Text View journal details
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