Developing genetic tools to exploit Chaetomium thermophilum for biochemical analyses of eukaryotic macromolecular assemblies

开发遗传工具利用嗜热毛壳菌进行真核大分子组装的生化分析

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作者:Nikola Kellner, Johannes Schwarz, Miriam Sturm, Javier Fernandez-Martinez, Sabine Griesel, Wenzhu Zhang, Brian T Chait, Michael P Rout, Ulrich Kück, Ed Hurt

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.

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