用于研究细胞转染的质粒DNA的序列特异性甲基转移酶诱导标记（SMILing）
Sequence-specific Methyltransferase-Induced Labelling (SMILing) of plasmid DNA for studying cell transfection.
作者信息
Schmidt Falk H-G, Hüben Michael, Gider Basar, Renault François, Teulade-Fichou Marie-Paule, Weinhold Elmar
机构信息
Laboratoire de Chimie des Interactions Moléculaires, Collège de France, Paris, France.
出版信息
Bioorg Med Chem. 2008 Jan 1;16(1):40-8. doi: 10.1016/j.bmc.2007.04.054. Epub 2007 May 3.
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.
使用新合成的N-腺苷氮丙啶辅因子和DNA甲基转移酶M.TaqI，将质粒DNA（pUC19和pBR322）进行序列特异性共价标记，标记上Cy3荧光团。将荧光标记的质粒用于转染哺乳动物细胞，并通过落射荧光显微镜和共聚焦荧光显微镜观察其在细胞内的分布。尽管这些原核质粒不含核输入序列，但仍观察到其向细胞核的转运。
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