Vector methylation inhibits transcription from the SV40 early promoter.
Methylation of a plasmid containing the SV40 promoter linked to the chloramphenicol acetyl transferase (CAT) gene, with either murine DNA methylase or methylase SssI results in inhibition of the expression of the reporter gene after transfection into cultured cells. Methylation of the plasmid with the methylases HhaI and HpaII has no effect on the expression of this gene. Protein-DNA interactions in the SV40 promoter are not affected by the presence of methylcytosine suggesting that inactivation results from the formation of an inactive chromatin structure that is dependent on the high CG content of the plasmid.
Bryans
M
M
Department of Biochemistry, University of Glasgow, UK.
Kass
S
S
Seivwright
C
C
Adams
R L
RL
D000818:Animals; D001483:Base Sequence; D002287:Carcinoma, Krebs 2 / Q000201:enzymology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D015254:DNA Modification Methylases / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D007700:Kinetics; D008745:Methylation; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection
D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D015254:DNA Modification Methylases; D015500:Chloramphenicol O-Acetyltransferase