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PMID: 6316255 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Replication and expression in mammalian cells of transfected DNA; description of an improved erythrocyte ghost fusion technique.

Nucleic acids research ·Vol. 11 ·No. 21 ·1983-11-11 ·Pages 7287-302

Wiberg FC, Sunnerhagen P, Kaltoft K, Zeuthen J, Bjursell G

Abstract

After modification of an erythrocyte - ghost fusion technique, DNA can be transferred in large amounts to mammalian cells. The overall recovery permits analysis of transferred DNA immediately after fusion by e.g. Southern - blot hybridization and electron microscopy. In general more than 20% of the cells contain the transfected DNA in their nuclei within 4 hours post fusion. Expression of Polyoma T antigens is delayed, as compared to a normal virus infection, and is detected at about 60 hours after transfer of DNA. During the first days an increase in amount of transferred DNA could be detected which might be due to limited replication. To analyze the replication of hybrid DNA molecules consisting of Polyoma and prokaryotic plasmid DNA sequences, fusion was followed by a normal Polyoma virus infection. The resulting induction of cells to enter the S phase made possible replication analysis of transfected hybrid DNA. Replicating molecules of both the theta form and rolling - circle type were observed.

MeSH Terms
Animals Cell Fusion Cell Line Cell Transformation, Viral Cells, Cultured Cloning, Molecular DNA Replication DNA, Viral/genetics Erythrocyte Membrane/metabolism Humans Mice Nucleic Acid Hybridization Polyomavirus/genetics Rats Transcription, Genetic Transfection
Chemicals
DNA, Viral
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Wiberg F C
Sunnerhagen P
Kaltoft K
Zeuthen J
Bjursell G
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29 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/11.21.7287
Published
1983-11-11
Pages
7287-302
Language
English
Region
England
NLM ID
0411011
PMCID
PMC326483
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/11.21.7287 PMC Full Text View journal details
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