Reference : Isolation of Nucleoli from Ehrlich Ascites Tumor Cells and Dynamics of Nascent RNA wi...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/30190
Isolation of Nucleoli from Ehrlich Ascites Tumor Cells and Dynamics of Nascent RNA within Isolated Nucleoli.
English
Thiry, Marc mailto [Université de Liège - ULg > Département des sciences de la vie > Biologie cellulaire >]
Ploton, Dominique [> > > >]
2008
Methods in Molecular Biology (Clifton, N.J.)
Humana Press
463
111-21
Yes (verified by ORBi)
International
1064-3745
Totowa
NJ
[en] Here we describe a new, rapid method for isolating nucleoli from Ehrlich tumor cells that preserves their morphological integrity and high transcriptional activity. Until now, methods for isolation of nucleoli were generally assumed to empty one of their three main compartments, the fibrillar center, of its contents. This new method consists of sonicating cells in an isotonic medium containing MgSO(4), spermidine, and spermine, followed by separation of nucleoli through a Percoll density gradient. Using the nonisotopic approach of labelling with BrUTP, we have further investigated the dynamics of nascent ribosomal RNAs (rRNAs) within morphologically intact isolated nucleoli at the electron microscope level. We show that ribosomal transcripts are elongated in the cortex of the fibrillar center and then enter the surrounding dense fibrillar component.
http://hdl.handle.net/2268/30190
10.1007/978-1-59745-406-3-8

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