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Proceedings Paper

Single-molecule fluorescence microscopy on nucleotide excision repair complexes using GFP fusion proteins
Author(s): Ine Segers-Nolten; Suzanne Rademakers; Wim Vermeulen; Aufried Lenferink; Cees Otto; Jan Hoeijmakers; Jan Greve
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Paper Abstract

Scanning Confocal Fluorescence Microscopy is used for single molecule studies on DNA-protein complexes that occur in Nucleotide Excision Repair (NER). During DNA-damage elimination by the NER-pathway, complex protein structures assemble over DNA. It is our aim to resolve the architecture of these DNA-protein complexes and to study dynamic changes that occur in these structures. For this purpose NER- complexes are partly reconstituted onto DNA-substrates using NER-proteins fused to different Green Fluorescent Protein mutants. Here we describe the recombinant production of NER- GFP fusion proteins. Characterization of GFP fluorescence is shown together with results of GFP single molecule imaging. First results with NER-GFP fusion proteins are presented as well.

Paper Details

Date Published: 18 December 2000
PDF: 9 pages
Proc. SPIE 4164, Laser Microscopy, (18 December 2000); doi: 10.1117/12.410629
Show Author Affiliations
Ine Segers-Nolten, Univ. of Twente (Netherlands)
Suzanne Rademakers, Erasmus Univ. Rotterdam (Netherlands)
Wim Vermeulen, Erasmus Univ. Rotterdam (Netherlands)
Aufried Lenferink, Univ. of Twente (Netherlands)
Cees Otto, Univ. of Twente (Netherlands)
Jan Hoeijmakers, Erasmus Univ. Rotterdam (Netherlands)
Jan Greve, Univ. of Twente (Netherlands)

Published in SPIE Proceedings Vol. 4164:
Laser Microscopy
Karsten Koenig; Hans J. Tanke; Herbert Schneckenburger, Editor(s)

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