title: Developing genetic tools to exploit Chaetomium thermophilum for biochemical analyses of eukaryotic macromolecular assemblies.
abstract: We describe a method to genetically manipulate Chaetomium thermophilum, a eukaryotic thermophile, along with various biochemical applications. The transformation method depends on a thermostable endogenous selection marker operating at high temperatures combined with chromosomal integration of target genes. Our technique allows exploiting eukaryotic thermophiles as source for purifying thermostable native macromolecular complexes with an emphasis on the nuclear pore complex, holding great potential for applications in basic science and biotechnology.
authors: Biochemistry Center, University of Heidelberg, Heidelberg, Germany.
authors: Nikola
authors: N
authors: Kellner
authors: Biochemistry Center, University of Heidelberg, Heidelberg, Germany.
authors: Johannes
authors: J
authors: Schwarz
authors: Biochemistry Center, University of Heidelberg, Heidelberg, Germany.
authors: Miriam
authors: M
authors: Sturm
authors: Laboratory of Cellular and Structural Biology and Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, New York, USA.
authors: Javier
authors: J
authors: Fernandez-Martinez
authors: Biochemistry Center, University of Heidelberg, Heidelberg, Germany.
authors: Sabine
authors: S
authors: Griesel
authors: Laboratory of Cellular and Structural Biology and Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, New York, USA.
authors: Wenzhu
authors: W
authors: Zhang
authors: Laboratory of Cellular and Structural Biology and Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, New York, USA.
authors: Brian T
authors: BT
authors: Chait
authors: Laboratory of Cellular and Structural Biology and Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, New York, USA.
authors: Michael P
authors: MP
authors: Rout
authors: Department for General and Molecular Botany, Ruhr-University Bochum, Bochum, Germany.
authors: Ulrich
authors: U
authors: Kück
authors: Biochemistry Center, University of Heidelberg, Heidelberg, Germany.
authors: Ed
authors: E
authors: Hurt
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology; D002597:Chaetomium / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D015825:Chromosomes, Fungal / Q000737:chemistry / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D006358:Hot Temperature; D009281:Naphthalenes / Q000494:pharmacology; D022022:Nuclear Pore / Q000235:genetics* / Q000378:metabolism; D028861:Nuclear Pore Complex Proteins / Q000235:genetics* / Q000378:metabolism; D011523:Protoplasts / Q000187:drug effects / Q000378:metabolism; D013172:Spores, Fungal / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D000077291:Terbinafine; D014170:Transformation, Genetic; D019076:Transgenes
chemical_list: D000935:Antifungal Agents; D005656:Fungal Proteins; D009281:Naphthalenes; D028861:Nuclear Pore Complex Proteins; D000077291:Terbinafine