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Dissertation zugänglich unter
URN: urn:nbn:de:gbv:18-23749
URL: http://ediss.sub.uni-hamburg.de/volltexte/2005/2374/

Sequence specific visualization of DNA in live mammalian cells

Mearini, Giulia

 Dokument 1.pdf (6.411 KB) 

Freie Schlagwörter (Englisch): chromatin dynamics, plasmid localization
Basisklassifikation: 42.13
Institut: Biologie
DDC-Sachgruppe: Biowissenschaften, Biologie
Dokumentart: Dissertation
Hauptberichter: Deppert, Wolfgang (Prof. Dr.)
Sprache: Englisch
Tag der mündlichen Prüfung: 10.12.2004
Erstellungsjahr: 2005
Publikationsdatum: 09.03.2005
Kurzfassung auf Englisch: In the first part of this work, the fate of plasmid DNA labeled with PNA was
investigated in living cells. It was shown that foreign DNA rapidly undergoes
interaction with intranuclear structural sites that strongly reduce its mobility
and restrict the DNA to regions excluding nucleoli and nuclear bodies as PML
bodies. The labeled plasmids partially colocalize with SAF-A, a well characterized
marker of the nuclear matrix, and are resistant towards extraction by detergent
and, in part, elevated salt concentration. Importantly the localization and low
mobility are independent of plasmid sequence and do not require the presence
of either a S/MAR DNA element or a functional promoter.
In the second part of this work, the attention was focused on chromatin
mobility in living mammalian cells by using the lac operator/repressor system.
The experiments show that the labeled chromatin loci are highly dynamic,
moving around by random walk, but are locally constrained in a defined nuclear
volume. Interestingly, the mobility of the analyzed loci depends on their location
and partially on the transcriptional status of the cell.


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