Sequence-specific Methyltransferase-Induced Labelling (SMILing) of plasmid DNA for studying cell transfection.
Plasmid DNA (pUC19 and pBR322) was sequence-specifically, covalently labelled with Cy3 fluorophores using a newly synthesised N-adenosylaziridine cofactor and the DNA methyltransferase M.TaqI. The fluorescently labelled plasmids were used for transfection of mammalian cells and their intracellular distribution was visualised by epifluorescence and confocal fluorescence microscopy. Although these prokaryotic plasmids do not contain nuclear import sequences, translocation into the nuclei was observed.
Schmidt
Falk H-G
FH
Laboratoire de Chimie des Interactions Moléculaires, Collège de France, Paris, France.
Hüben
Michael
M
Gider
Basar
B
Renault
François
F
Teulade-Fichou
Marie-Paule
MP
Weinhold
Elmar
E
D021581:Active Transport, Cell Nucleus; D000818:Animals; D001483:Base Sequence; D002232:Carbocyanines; D002478:Cells, Cultured; D015254:DNA Modification Methylases; D006801:Humans; D015336:Molecular Probe Techniques; D010957:Plasmids; D019914:Taq Polymerase; D014162:Transfection
D002232:Carbocyanines; C089187:cyanine dye 3; D015254:DNA Modification Methylases; D019914:Taq Polymerase