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Polyethylene Glycol-mediated Transformation of Drechmeria coniospora.

Bio-protocol (2017-03-05)
Le D He, Jonathan J Ewbank
ABSTRACT

Drechmeria coniospora is a nematophagous fungus and potential biocontrol agent. It belongs to the Ascomycota. It is related to Hirsutella minnesotensis, another nematophagous fungus but, phylogenetically, it is currently closest to the truffle parasite Tolypocladium ophioglossoides. Together with its natural host, Caenorhabditis elegans, it is used to study host-pathogen interactions. Here, we report a polyethylene glycol-mediated transformation method ( Turgeon et al., 2010 ; Ochman et al., 1988 ) for this fungus. The protocol can be used to generate both knock-in or knock-out strains ( Lebrigand et al., 2016 ).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Trizma® hydrochloride, BioXtra, pH 3.5-5.0 (0.5 M in H2O), ≥99.0% (titration)
Sigma-Aldrich
Cholesterol, powder, BioReagent, suitable for cell culture, ≥99%
Sigma-Aldrich
Aurintricarboxylic Acid, A cell-permeable polyanionic, polyaromatic compound used as a powerful inhibitor of cellular processes that are dependent on the formation of protein-nucleic acid complexes.
Sigma-Aldrich
Gentamicin solution, 10 mg/mL in deionized water, liquid, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Ampicillin sodium salt
Sigma-Aldrich
Magnesium sulfate, anhydrous, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Calcium chloride, anhydrous, granular, ≤7.0 mm, ≥93.0%