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End-joining of reconstituted histone H2AX-containing chromatin in vitro by soluble nuclear proteins from human cells
Authors:Siino Joseph S  Nazarov Igor B  Zalenskaya Irina A  Yau Peter M  Bradbury E Morton  Tomilin Nikolai V
Institution:Max-Planck-Institute for Infection Biology, Department of Molecular Biology, Schumannstr. 21/22, D-10117 Berlin, Germany.
Abstract:Green fluorescent protein (GFP) is an attractive reporter for Salmonella gene expression analysis but might interfere with virulence when expressed at high levels. To identify suitable GFP levels, we constructed a series of Salmonella strains expressing different amounts of GFP and measured their fluorescence and colonization levels in infected mice. The results show that GFP concentrations in the range of 7000-200,000 molecules per Salmonella cell are detectable in ex vivo samples using flow cytometry, and cause no major Salmonella virulence defect. Appropriate GFP levels can be obtained with weak promoters and stable GFP, or strong promoters and destabilized GFP.
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