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PMID: 3859838 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Minichromosome assembly of non-integrated plasmid DNA transfected into mammalian cells.

Nucleic acids research ·Vol. 13 ·No. 10 ·1985-05-24 ·Pages 3599-615

Reeves R, Gorman CM, Howard B

Abstract

The nucleoprotein structures formed on various plasmid expression vectors transfected into mammalian cells by both the calcium phosphate and DEAE-dextran methods have been studied. We demonstrate by a variety of means that mammalian cells are capable of rapidly assembling non-integrated circular plasmids (both replicating and non-replicating) into typical "minichromosomes" containing nucleosomes with a 190 bp repetitive spacing. Treatment of recipient cells with sodium butyrate for a short period of time (12-16 h) immediately following transfection markedly increased the DNase I digestion sensitivity of the newly assembled plasmid chromatin. Furthermore, minichromosomes isolated from such butyrate-treated cells are depleted in histone H1 and contain highly acetylated forms of histone H4. These findings are entirely consistent with our earlier speculation (Gorman et al., Nucleic Acids Res. 11, 1044; 1983) that appropriate butyrate treatment might stimulate transient expression of newly transfected genes by facilitating their assembly into an "active" type of chromatin structure.

MeSH Terms
Acetylation Acetyltransferases/genetics Animals Butyrates/pharmacology Butyric Acid Calcium Phosphates/pharmacology Cells, Cultured Chloramphenicol O-Acetyltransferase Chlorocebus aethiops Chromatin/analysis Chromosomes DEAE-Dextran/pharmacology DNA Replication Gene Expression Regulation Histones/metabolism Mice Plasmids Transfection
Chemicals
Butyrates Calcium Phosphates Chromatin Histones alpha-tricalcium phosphate tetracalcium phosphate Butyric Acid calcium phosphate, monobasic, anhydrous DEAE-Dextran calcium phosphate Acetyltransferases Chloramphenicol O-Acetyltransferase calcium phosphate, dibasic, anhydrous
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Reeves R
Gorman C M
Howard B
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32 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/13.10.3599
Published
1985-05-24
Pages
3599-615
Language
English
Region
England
NLM ID
0411011
PMCID
PMC341261
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/13.10.3599 PMC Full Text View journal details
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