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. 2011 Jan 11;30(3):617–626. doi: 10.1038/emboj.2010.345

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Copyright © 2011, European Molecular Biology Organization

This is an open-access article distributed under the terms of the Creative Commons Attribution Noncommercial Share Alike 3.0 Unported License, which allows readers to alter, transform, or build upon the article and then distribute the resulting work under the same or similar license to this one. The work must be attributed back to the original author and commercial use is not permitted without specific permission.

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Characteristics of a SepF mutant defective in ring formation. (A) SepF–FtsZ co-elution experiment using different MBP–SepF mutants. The elution fractions where analyzed by SDS–PAGE and Coomassie staining. The columns were incubated with (+) or without (−) B. subtilis FtsZ. As a negative control A98V and F124S were included. (B) Localization of SepF–G135N–GFP fusion in ΔsepF B. subtilis cells. The fusion protein (white bands) is located at cell division sites. (C) Images of negatively stained purified SepF–G135N (left panel), and in the presence of FtsZ (right panel, FtsZ protofilaments are clearly visible). Scale bar: 200 nm. (D) Mutation G135N is unable to sustain growth in the absence of FtsA. (E) Western blot analysis of SepF levels in wild-type B. subtilis cells (wt), a sepF knockout strain (ΔsepF), and a strain expressing SepF–G135N (G135N).