Academic
Publications
Visualization of RecA Filaments and DNA by Fluorescence Microscopy

Visualization of RecA Filaments and DNA by Fluorescence Microscopy,Taro Nishinaka,Yuko Doi,Makiko Hashimoto,Reiko Hara,Takehiko Shibata,Yoshie Harada,

Visualization of RecA Filaments and DNA by Fluorescence Microscopy   (Citations: 3)
BibTex | RIS | RefWorks Download
We have developed two experimental methods for observing Escherichia coli RecA-DNA filament under a fluorescence microscope. First, RecA-DNA filaments were visualized by immunofluorescence staining with anti-RecA monoclonal anti- body. Although the detailed filament structures below submicron scale were unable to be measured accurately due to optical resolution limit, this method has an advantage to analyse a large number of RecA-DNA filaments in a single experiment. Thus, it provides a reliable statistical distribution of the filament morphology. Moreover, not only RecA filament, but also naked DNA region was visualized separately in combination with immunofluorescence staining using anti-DNA monoclonal antibody. Second, by using cysteine derivative RecA protein, RecA-DNA filament was directly labelled by fluorescent reagent, and was able to observe directly under a fluorescence microscope with its enzymatic activity maintained. We showed that the RecA-DNA filament disassembled in the direction from 50 to 30 of ssDNA as dATP hydrolysis proceeded.
Published in 2007.
Cumulative Annual
Sort by: