Fur homodimerization and functional assays

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Presentation transcript:

Fur homodimerization and functional assays Fur homodimerization and functional assays.A, yeast two-hybrid competition assay. Fur homodimerization and functional assays.A, yeast two-hybrid competition assay. We co-expressed in yeast LexA-Fur, NLS-B42-HA-Fur, and different HA-NLS-aptamer or -Fur fusions. We quantified the Fur dimerization two-hybrid phenotypes using a luciferase reporter gene, and we normalized the results with the value obtained with aptamer cl20. We performed three independent experiments. B, bacterial growth on medium containing a high concentration of Mn2+. We spotted serial dilutions of E. coli XL-1 blue transformants expressing peptide aptamers or Fur90 onto TN medium containing 10 mm Mn2+ and arabinose. C, Fur transcriptional repression assay. We transformed QC6008 (fur−) strain with pBAD24 to confer ampicillin resistance and QC2146 (fur+) strain with pBAD24 expressing peptide aptamers, Fur90, or nothing. We streaked the transformants onto solid medium containing X-gal with or without arabinose. D, oxidative stress assay. We measured the growth of E. coli QC2949 (fur+) transformants expressing various aptamers or Fur90 and the growth of QC6009 (fur−) transformed with pBAD24 in the presence or in the absence of H2O2. We performed four independent experiments. Nadia Abed et al. Mol Cell Proteomics 2007;6:2110-2121 © 2007 The American Society for Biochemistry and Molecular Biology