SPINE: a method for the rapid detection and analysis of protein-protein interactions in vivo |
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Authors: | Herzberg Christina Weidinger Lope Andrés Flórez Dörrbecker Bastian Hübner Sebastian Stülke Jörg Commichau Fabian M |
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Institution: | 1. Department of General Microbiology, Georg‐August‐University G?ttingen, G?ttingen, Germany;2. Department of General Microbiology, Georg‐August‐University G?ttingen, G?ttingen, GermanyDepartment of General Microbiology, Georg‐August‐University G?ttingen, Grisebachstr. 8, 37077 G?ttingen, Germany Fax: +49‐551‐393808=== |
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Abstract: | The detection and analysis of protein-protein interactions is one of the central tasks of proteomics in the postgenomic era. For this purpose, we present a procedure, the Strep-protein interaction experiment (SPINE) that combines the advantages of the Strep-tag protein purification system with those of reversible in vivo protein crosslinking by formaldehyde. Using two Bacillus subtilis regulator proteins, we demonstrate that this method is well suited to isolate protein complexes with high purity and virtually no background. Plasmids allowing the high-level expression of proteins carrying an N- or C-terminal Strep-tag in B. subtilis were constructed. |
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Keywords: | Bacillus subtilis Crosslinking Protein–protein interaction SPINE Strep‐tag |
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