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  • Conditional Knockdown of Proteins Using Auxin-inducible Degron (AID) Fusions in Toxoplasma gondii.

Conditional Knockdown of Proteins Using Auxin-inducible Degron (AID) Fusions in Toxoplasma gondii.

Bio-protocol (2018-04-13)
Kevin M Brown, Shaojun Long, L David Sibley
要旨

Toxoplasma gondii is a member of the deadly phylum of protozoan parasites called Apicomplexa. As a model apicomplexan, there is a great wealth of information regarding T. gondii's 8,000+ protein coding genes including sequence variation, expression, and relative contribution to parasite fitness. However, new tools are needed to functionally investigate hundreds of putative essential protein coding genes. Accordingly, we recently implemented the auxin-inducible degron (AID) system for studying essential proteins in T. gondii. Here we provide a step-by-step protocol for examining protein function in T. gondii using the AID system in a tissue culture setting.

材料
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製品内容

Sigma-Aldrich
リン酸カリウム 一塩基性, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
Sigma-Aldrich
TWEEN® 20, viscous liquid, suitable for cell culture
Sigma-Aldrich
L-グルタチオン、還元型, suitable for cell culture, BioReagent, ≥98.0%, powder
Sigma-Aldrich
ホウ酸, BioReagent, for molecular biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%
Sigma-Aldrich
リン酸ナトリウム 二塩基性, for molecular biology, ≥98.5% (titration)
Sigma-Aldrich
アンピシリン ナトリウム塩
Sigma-Aldrich
ドデシル硫酸ナトリウム, 92.5-100.5% based on total alkyl sulfate content basis
Sigma-Aldrich
塩化カルシウム 二水和物, BioXtra, ≥99.0%