在……中使用生长素诱导降解结构域（AID）融合蛋白进行蛋白质的条件性敲低
Conditional Knockdown of Proteins Using Auxin-inducible Degron (AID) Fusions in .
作者信息
Brown Kevin M, Long Shaojun, Sibley L David
机构信息
Department of Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri, USA.
出版信息
Bio Protoc. 2018 Feb 20;8(4). doi: 10.21769/BioProtoc.2728.
Abstract
is a member of the deadly phylum of protozoan parasites called Apicomplexa. As a model apicomplexan, there is a great wealth of information regarding '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 . Here we provide a step-by-step protocol for examining protein function in using the AID system in a tissue culture setting.
摘要
是被称为顶复门的致命原生动物寄生虫门的成员。作为一种典型的顶复门生物，关于其8000多个蛋白质编码基因有大量信息，包括序列变异、表达以及对寄生虫适应性的相对贡献。然而，需要新的工具来对数百个推定的必需蛋白质编码基因进行功能研究。因此，我们最近实施了生长素诱导降解子（AID）系统来研究中的必需蛋白质。在这里，我们提供了一个在组织培养环境中使用AID系统研究中蛋白质功能的逐步方案。
相似文献
1
Conditional Knockdown of Proteins Using Auxin-inducible Degron (AID) Fusions in .在……中使用生长素诱导降解结构域（AID）融合蛋白进行蛋白质的条件性敲低
Bio Protoc. 2018 Feb 20;8(4). doi: 10.21769/BioProtoc.2728.
2
Prenyl Transferases Regulate Secretory Protein Sorting and Parasite Morphology in . prenyl 转移酶调节. 中的分泌蛋白分拣和寄生虫形态
Int J Mol Sci. 2023 Apr 12;24(8):7172. doi: 10.3390/ijms24087172.
3
Coupling Auxin-Inducible Degron System with Ultrastructure Expansion Microscopy to Accelerate the Discovery of Gene Function in Toxoplasma gondii.将生长素诱导降解系统与超微结构扩展显微镜相结合，加速弓形虫基因功能的发现。
Methods Mol Biol. 2021;2369:121-137. doi: 10.1007/978-1-0716-1681-9_8.
4
Plasma Membrane Association by N-Acylation Governs PKG Function in .通过N-酰化作用实现的质膜关联调控了……中的蛋白激酶G功能
mBio. 2017 May 2;8(3):e00375-17. doi: 10.1128/mBio.00375-17.
5
The plant-like protein phosphatase PPKL regulates parasite replication and morphology in Toxoplasma gondii.植物样蛋白磷酸酶 PPKL 调控弓形虫中的寄生虫复制和形态。
Parasit Vectors. 2024 Mar 18;17(1):142. doi: 10.1186/s13071-024-06135-6.
6
AID-ing Signaling in .艾滋病相关信号传导于…… （你提供的原文“AID-ing Signaling in.”似乎不完整，翻译可能不太准确，你可以补充完整后再让我翻译）
mBio. 2017 Jul 25;8(4):e01076-17. doi: 10.1128/mBio.01076-17.
7
CRISPR/Cas9-Mediated Generation of Tetracycline Repressor-Based Inducible Knockdown in Toxoplasma gondii.CRISPR/Cas9 介导的基于四环素抑制物的诱导性敲低在刚地弓形虫中的产生。
Methods Mol Biol. 2020;2071:125-141. doi: 10.1007/978-1-4939-9857-9_7.
8
Hypothetical bromodomain-containing protein 5 is required for the growth of Toxoplasma gondii.假定的含溴结构域蛋白 5 是刚地弓形虫生长所必需的。
Vet Parasitol. 2022 Sep;309:109767. doi: 10.1016/j.vetpar.2022.109767. Epub 2022 Jul 22.
9
Characterisation of the OTU domain deubiquitinase complement of .OTU 结构域去泛素化酶复合体的特性研究。
Life Sci Alliance. 2023 Mar 23;6(6). doi: 10.26508/lsa.202201710. Print 2023 Jun.
10
Transcriptional Profiling Suggests T Cells Cluster around Neurons Injected with Toxoplasma gondii Proteins.转录谱分析表明，T 细胞聚集在注射弓形虫蛋白的神经元周围。
mSphere. 2020 Sep 2;5(5):e00538-20. doi: 10.1128/mSphere.00538-20.
引用本文的文献
1
The PP2A-2 holoenzyme orchestrates daughter cell emergence during cytokinesis in Toxoplasma gondii.PP2A-2全酶在弓形虫胞质分裂过程中协调子细胞的出现。
PLoS Pathog. 2025 Sep 3;21(9):e1013475. doi: 10.1371/journal.ppat.1013475. eCollection 2025 Sep.
2
AP2X-1 is a negative regulator of sexual commitment.AP2X-1是性成熟的负调控因子。
mBio. 2025 Sep 10;16(9):e0005225. doi: 10.1128/mbio.00052-25. Epub 2025 Aug 18.
3
Co-dependent formation of the subpellicular microtubules and inner membrane skeleton.表膜下微管和内膜骨架的共依赖性形成。
mBio. 2025 Aug 13:e0138925. doi: 10.1128/mbio.01389-25.
4
A rapid, simple, and economical method for the isolation of ribosomes and translational machinery for structural and functional studies.一种用于分离核糖体及翻译机制以进行结构和功能研究的快速、简便且经济的方法。
Nat Commun. 2025 Aug 5;16(1):7185. doi: 10.1038/s41467-025-62314-8.
5
Deciphering cell cycle organization of endodyogeny.解析内二均裂的细胞周期组织
mBio. 2025 Jul 1:e0111925. doi: 10.1128/mbio.01119-25.
6
Sarcoendoplasmic reticulum calcium ATPase is an essential and druggable lipid-dependent ion pump in Toxoplasma gondii.肌质内质网钙ATP酶是刚地弓形虫中一种重要的、可药物作用的脂质依赖性离子泵。
Commun Biol. 2025 May 6;8(1):702. doi: 10.1038/s42003-025-08058-z.
7
The essential kinase TgGSK regulates centrosome segregation and endodyogeny in .必需激酶TgGSK调节……中的中心体分离和内二分裂。 （原文句子不完整，缺少具体的研究对象）
mSphere. 2025 Apr 29;10(4):e0011125. doi: 10.1128/msphere.00111-25. Epub 2025 Mar 28.
8
PROP1 is critical for autophagy and parasite viability during chronic infection.PROP1在慢性感染期间对自噬和寄生虫的生存能力至关重要。
mSphere. 2025 Mar 25;10(3):e0082924. doi: 10.1128/msphere.00829-24. Epub 2025 Feb 21.
9
Phospho-relay feedback loops control egress vs. intracellular development in Toxoplasma gondii.磷酸化中继反馈环控制弓形虫的出芽与细胞内发育。
Cell Rep. 2025 Feb 25;44(2):115260. doi: 10.1016/j.celrep.2025.115260. Epub 2025 Feb 3.
10
TgATG9 is required for autophagosome biogenesis and maintenance of chronic infection in .TgATG9对于自噬体的生物发生以及疟原虫慢性感染的维持是必需的。 （注：原英文文本不完整，这里补充了“疟原虫”使句子完整通顺，不然单看原英文不知道是在说什么生物的感染等情况）
Autophagy Rep. 2024;3(1). doi: 10.1080/27694127.2024.2418256. Epub 2024 Oct 23.
本文引用的文献
1
A conserved ankyrin repeat-containing protein regulates conoid stability, motility and cell invasion in Toxoplasma gondii.一种保守的含锚蛋白重复序列的蛋白调节刚毛稳定性、运动性和细胞侵袭在弓形体。
Nat Commun. 2017 Dec 21;8(1):2236. doi: 10.1038/s41467-017-02341-2.
2
Calmodulin-like proteins localized to the conoid regulate motility and cell invasion by Toxoplasma gondii.定位于类锥体的类钙调蛋白调节刚地弓形虫的运动性和细胞侵袭。
PLoS Pathog. 2017 May 5;13(5):e1006379. doi: 10.1371/journal.ppat.1006379. eCollection 2017 May.
3
Plasma Membrane Association by N-Acylation Governs PKG Function in .通过N-酰化作用实现的质膜关联调控了……中的蛋白激酶G功能
mBio. 2017 May 2;8(3):e00375-17. doi: 10.1128/mBio.00375-17.
4
Toxoplasma gondii plaque assays revisited: Improvements for ultrastructural and quantitative evaluation of lytic parasite growth.弓形虫空斑试验再探讨：溶细胞性寄生虫生长的超微结构及定量评估的改进
Exp Parasitol. 2017 Sep;180:19-26. doi: 10.1016/j.exppara.2016.12.015. Epub 2016 Dec 21.
5
Development of CRISPR/Cas9 for Efficient Genome Editing in Toxoplasma gondii.用于在弓形虫中进行高效基因组编辑的CRISPR/Cas9技术的开发。
Methods Mol Biol. 2017;1498:79-103. doi: 10.1007/978-1-4939-6472-7_6.
6
Conditional Degradation of Plasmodium Calcineurin Reveals Functions in Parasite Colonization of both Host and Vector.疟原虫钙调神经磷酸酶的条件性降解揭示其在宿主和媒介寄生虫定殖中的功能
Cell Host Microbe. 2015 Jul 8;18(1):122-31. doi: 10.1016/j.chom.2015.05.018. Epub 2015 Jun 25.
7
Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation.用于CRISPR-Cas9介导的基因失活的高活性sgRNA的合理设计。
Nat Biotechnol. 2014 Dec;32(12):1262-7. doi: 10.1038/nbt.3026. Epub 2014 Sep 3.
8
Efficient genome engineering of Toxoplasma gondii using CRISPR/Cas9.利用CRISPR/Cas9对刚地弓形虫进行高效基因组工程改造。
PLoS One. 2014 Jun 27;9(6):e100450. doi: 10.1371/journal.pone.0100450. eCollection 2014.
9
Efficient gene disruption in diverse strains of Toxoplasma gondii using CRISPR/CAS9.利用CRISPR/CAS9在多种弓形虫菌株中实现高效基因破坏。
mBio. 2014 May 13;5(3):e01114-14. doi: 10.1128/mBio.01114-14.
10
2A and the auxin-based degron system facilitate control of protein levels in Plasmodium falciparum.2A 元件和基于生长素的降解结构域系统有助于控制疟原虫中的蛋白质水平。
PLoS One. 2013 Nov 13;8(11):e78661. doi: 10.1371/journal.pone.0078661. eCollection 2013.
Zotero 插件