Craige Branch, Brown Jason M, Witman George B
Department of Cell and Developmental Biology, University of Massachusetts Medical School, Worcester, Massachusetts.
Curr Protoc Cell Biol. 2013 Jun;Chapter 3:3.41.1-3.41.9. doi: 10.1002/0471143030.cb0341s59.
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.
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。
