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
复制标题
DOI：
10.21769/bioprotoc.2728
复制
发表时间：
2018-02-20
期刊：
影响因子：
0.8
通讯作者：
Sibley, L. David
中科院分区：
文献类型：
--
作者：
Brown, Kevin M.;Long, Shaojun;Sibley, L. David
弓形虫是一种致命的原生动物寄生虫门的成员，称为顶复门。作为一种典型的顶复形动物，关于T.弓形虫的8,000多个蛋白质编码基因，包括序列变异，表达和对寄生虫适应性的相对贡献。然而，需要新的工具来功能性地研究数百个推定的必需蛋白质编码基因。因此，我们最近实施了生长素诱导降解决定子（AID）系统来研究T。刚地。在这里，我们提供了一个一步一步的协议，检查蛋白质的功能，在T。在组织培养环境中使用AID系统培养弓形虫。
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.