衣藻鞭毛的分离
Isolation of Chlamydomonas flagella.
作者信息
Craige Branch, Brown Jason M, Witman George B
机构信息
Department of Cell and Developmental Biology, University of Massachusetts Medical School, Worcester, Massachusetts.
出版信息
Curr Protoc Cell Biol. 2013 Jun;Chapter 3:3.41.1-3.41.9. doi: 10.1002/0471143030.cb0341s59.
Abstract
A simple, scalable, and fast procedure for the isolation of Chlamydomonas flagella is described. Chlamydomonas can be synchronously deflagellated by treatment with chemicals, pH shock, or mechanical shear. The Basic Protocol describes the procedure for flagellar isolation using dibucaine to induce flagellar abscission; we also describe the pH shock method as an Alternate Protocol when flagellar regeneration is desirable. Sub-fractionation of the isolated flagella into axonemes and the membrane + matrix fraction is described in a Support Protocol.
摘要
本文描述了一种简单、可扩展且快速的分离衣藻鞭毛的方法。衣藻可以通过化学处理、pH 冲击或机械剪切实现同步去鞭毛。基本方案描述了使用丁卡因诱导鞭毛脱落来分离鞭毛的步骤；当需要鞭毛再生时，我们还将 pH 冲击法作为替代方案进行了描述。在支持方案中描述了将分离的鞭毛进一步亚分级为轴丝以及膜 + 基质部分的方法。
相似文献
1
Isolation of Chlamydomonas flagella.衣藻鞭毛的分离
Curr Protoc Cell Biol. 2013 Jun;Chapter 3:3.41.1-3.41.9. doi: 10.1002/0471143030.cb0341s59.
2
Isolation of Chlamydomonas flagella and flagellar axonemes.衣藻鞭毛和鞭毛轴丝的分离
Methods Enzymol. 1986;134:280-90. doi: 10.1016/0076-6879(86)34096-5.
3
Isolation of radial spoke heads from Chlamydomonas axonemes.
Methods Cell Biol. 1995;47:381-3. doi: 10.1016/s0091-679x(08)60833-x.
4
Calcium control of waveform in isolated flagellar axonemes of Chlamydomonas.衣藻分离鞭毛轴丝中波形的钙调控
J Cell Biol. 1980 Aug;86(2):446-55. doi: 10.1083/jcb.86.2.446.
5
Flagellar coordination in Chlamydomonas reinhardtii: isolation and reactivation of the flagellar apparatus.莱茵衣藻的鞭毛协调作用：鞭毛装置的分离与重新激活
Science. 1975 Sep 12;189(4206):891-3. doi: 10.1126/science.1098148.
6
Analysis of force generation during flagellar assembly through optical trapping of free-swimming Chlamydomonas reinhardtii.通过光学捕获自由游动的莱茵衣藻分析鞭毛组装过程中的力产生
Cell Motil Cytoskeleton. 2005 Jul;61(3):137-44. doi: 10.1002/cm.20071.
7
Comparative structural analysis of eukaryotic flagella and cilia from Chlamydomonas, Tetrahymena, and sea urchins.真核生物鞭毛和纤毛的比较结构分析：来自衣藻、四膜虫和海胆。
J Struct Biol. 2012 May;178(2):199-206. doi: 10.1016/j.jsb.2012.02.012. Epub 2012 Mar 3.
8
Centrin-mediated microtubule severing during flagellar excision in Chlamydomonas reinhardtii.莱茵衣藻鞭毛切除过程中中心蛋白介导的微管切断
J Cell Biol. 1989 May;108(5):1751-60. doi: 10.1083/jcb.108.5.1751.
9
Large-scale isolation of Chlamydomonas flagella.衣藻鞭毛的大规模分离
Methods Cell Biol. 1995;47:9-12. doi: 10.1016/s0091-679x(08)60783-9.
10
A mutant of Chlamydomonas reinhardtii that lacks the flagellar outer dynein arm but can swim.莱茵衣藻的一种突变体，缺乏鞭毛外动力蛋白臂但仍能游动。
J Cell Sci. 1985 Mar;74:181-91. doi: 10.1242/jcs.74.1.181.
引用本文的文献
1
A conserved mechanism for the retrieval of polyubiquitinated proteins from cilia.一种从纤毛中回收多聚泛素化蛋白的保守机制。
Cell. 2025 Aug 18. doi: 10.1016/j.cell.2025.07.043.
2
Snow algae exhibit diverse motile behaviors and thermal responses.雪藻表现出多样的运动行为和热反应。
mBio. 2025 May 14;16(5):e0295424. doi: 10.1128/mbio.02954-24. Epub 2025 Apr 1.
3
Tubulin tyrosination/detyrosination regulate the affinity and sorting of intraflagellar transport trains on axonemal microtubule doublets.微管蛋白酪氨酸化/去酪氨酸化调节轴丝微管双联体上鞭毛内运输列车的亲和力和分选。
Nat Commun. 2025 Jan 26;16(1):1055. doi: 10.1038/s41467-025-56098-0.
4
IC97, an intermediate chain of the flagellar dynein f/I1, is required for normal flagellar and cellular motility.IC97是鞭毛动力蛋白f/I1的中间链，是正常鞭毛和细胞运动所必需的。
mSphere. 2024 Dec 19;9(12):e0055824. doi: 10.1128/msphere.00558-24. Epub 2024 Nov 27.
5
Transcriptional analysis of primary ciliary dyskinesia airway cells reveals a dedicated cilia glutathione pathway.原发性纤毛运动障碍气道细胞的转录组分析揭示了一种特有的纤毛谷胱甘肽途径。
JCI Insight. 2024 Jul 23;9(17):e180198. doi: 10.1172/jci.insight.180198.
6
Mastigoneme structure reveals insights into the O-linked glycosylation code of native hydroxyproline-rich helices.Mastigoneme 结构揭示了对天然羟脯氨酸丰富螺旋 O-连接糖基化密码的深入了解。
Cell. 2024 Apr 11;187(8):1907-1921.e16. doi: 10.1016/j.cell.2024.03.005. Epub 2024 Mar 28.
7
Gene dosage of independent dynein arm motor preassembly factors influences cilia assembly in Chlamydomonas reinhardtii.独立的动力蛋白臂马达预组装因子的基因剂量影响莱茵衣藻纤毛的组装。
PLoS Genet. 2024 Mar 18;20(3):e1011038. doi: 10.1371/journal.pgen.1011038. eCollection 2024 Mar.
8
Structural determination and modeling of ciliary microtubules.纤毛微管的结构测定和建模。
Acta Crystallogr D Struct Biol. 2024 Apr 1;80(Pt 4):220-231. doi: 10.1107/S2059798324001815. Epub 2024 Mar 7.
9
The Small Interactor of PKD2 protein promotes the assembly and ciliary entry of the Chlamydomonas PKD2-mastigoneme complexes.PKD2 蛋白的小相互作用蛋白促进了衣藻 PKD2-纤毛蛋白复合物的组装和纤毛进入。
J Cell Sci. 2024 Jan 1;137(1). doi: 10.1242/jcs.261497. Epub 2024 Jan 12.
10
Cryo-EM reveals how the mastigoneme assembles and responds to environmental signal changes.冷冻电镜揭示了mastigoneme 如何组装以及如何响应环境信号变化。
J Cell Biol. 2023 Dec 4;222(12). doi: 10.1083/jcb.202301066. Epub 2023 Oct 26.
Zotero 插件