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Polyethylene Glycol-mediated Transformation of Drechmeria coniospora.

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

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 ).

材料
製品番号
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製品内容

Sigma-Aldrich
硫酸マグネシウム, anhydrous, ReagentPlus®, ≥99.5%
Sigma-Aldrich
塩化カルシウム, anhydrous, granular, ≤7.0 mm, ≥93.0%
Sigma-Aldrich
ゲンタマイシン 溶液, 10 mg/mL in deionized water, liquid, 0.1 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
コレステロール, powder, BioReagent, suitable for cell culture, ≥99%
Sigma-Aldrich
Trizma® 塩酸塩, BioXtra, pH 3.5-5.0 (0.5 M in H2O), ≥99.0% (titration)
Sigma-Aldrich
アンピシリン ナトリウム塩
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.