Single-molecule fluorescence microscopy on nucleotide excision repair complexes using GFP fusion proteins

Ine Segers-Nolten; Suzanne Rademakers; Wim Vermeulen; Aufried Lenferink; Cees Otto; Jan Hoeijmakers; Jan Greve

doi:10.1117/12.410629

18 December 2000 Single-molecule fluorescence microscopy on nucleotide excision repair complexes using GFP fusion proteins

Ine Segers-Nolten, Suzanne Rademakers, Wim Vermeulen, Aufried Lenferink, Cees Otto, Jan Hoeijmakers, Jan Greve

Author Affiliations +

Proceedings Volume 4164, Laser Microscopy; (2000) https://doi.org/10.1117/12.410629
Event: EOS/SPIE European Biomedical Optics Week, 2000, Amsterdam, Netherlands

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.

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Ine Segers-Nolten, Suzanne Rademakers, Wim Vermeulen, Aufried Lenferink, Cees Otto, Jan Hoeijmakers, and Jan Greve "Single-molecule fluorescence microscopy on nucleotide excision repair complexes using GFP fusion proteins", Proc. SPIE 4164, Laser Microscopy, (18 December 2000); https://doi.org/10.1117/12.410629

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KEYWORDS

Proteins

Molecules

Green fluorescent protein

Luminescence

Confocal microscopy

Microscopy

Diffusion

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