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Merck

Polyethylene Glycol-mediated Transformation of Drechmeria coniospora.

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

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 ).

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Sigma-Aldrich
硫酸镁, anhydrous, ReagentPlus®, ≥99.5%
Sigma-Aldrich
氯化钙, anhydrous, granular, ≤7.0 mm, ≥93.0%
Sigma-Aldrich
庆大霉素 溶液, 10 mg/mL in deionized water, liquid, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
胆固醇, powder, BioReagent, suitable for cell culture, ≥99%
Sigma-Aldrich
氨苄西林 钠盐
Sigma-Aldrich
Trizma® 盐酸盐 盐酸盐, BioXtra, pH 3.5-5.0 (0.5 M in H2O), ≥99.0% (titration)
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.