Transformation of Chaetomium thermophilum and Affinity Purification of Native Thermostable Protein Complexes

Methods Mol Biol. 2022:2502:35-50. doi: 10.1007/978-1-0716-2337-4_2.

Abstract

Chaetomium thermophilum, a eukaryotic thermophile, is an aspiring organism holding great potential for various biochemical and biotechnological applications. Prerequisite for genetic manipulation is a reliable transformation system for target genes combined with selection markers operating at the high growth temperatures of the fungus. Here, we present a detailed protocol for Chaetomium thermophilum protoplast transformation to allow stable chromosomal integration of constructs into its genome, rendering this eukaryotic thermophile a valuable resource for affinity purification of native thermostable protein complexes, like nuclear pore subcomplexes.

Keywords: Affinity purification; Chaetomium thermophilum; Genetic transformation; Nuclear pore complex (NPC); Thermophile.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chaetomium* / genetics
  • Chaetomium* / metabolism
  • Chromatography, Affinity
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Nuclear Pore / metabolism
  • Nuclear Pore Complex Proteins / metabolism

Substances

  • Fungal Proteins
  • Nuclear Pore Complex Proteins

Supplementary concepts

  • Chaetomium thermophilum