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Methods Mol Biol. 2012;944:191-205. doi: 10.1007/978-1-62703-122-6_14.

Identification of protein complexes from filamentous fungi with tandem affinity purification.

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  • 1Abteilung Molekulare Mikrobiologie und Genetik, DFG Research Center for Molecular Physiology of the Brain (CMPB), Georg-August-Universität Göttingen, Göttingen, Germany.

Abstract

Fungal molecular biology has benefited from the enormous advances in understanding protein-protein interactions in prokaryotic or eukaryotic organisms of the past decade. Tandem affinity purification (TAP) allows the enrichment of native protein complexes from cell extracts under mild conditions. We codon-optimized tags and established TAP, previously not applicable to filamentous fungi, for the model organism Aspergillus nidulans. We could identify by this method the trimeric Velvet complex VelB/VeA/LaeA or the eight subunit COP9 signalosome. Here, we describe an optimized protocol for A. nidulans which can also be adapted to other filamentous fungi.

PMID:
23065618
[PubMed - indexed for MEDLINE]

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