CIL:13881

Tem1-CAAX (green) is targeted away from spindle pole bodies in wild-type cells (CIL# 13880) but can also still localize to the SPB (this image). The CAAX motif from Ras2 targets proteins to the cytoplasmic side of the membrane. tem1Δ::GAL-UPL-TEM1 cells expressing eGFP-TEM1-CAAX from a CEN plasmid were grown on 2% raffinose/2% galactose and transferred to 2% glucose medium prior to image capture. A differential interference contrast (DIC) image is also shown. The tem1Δ::GAL1-UPL-TEM1 strain allows for the rapid, conditional depletion of Tem1. UPL, which stands for ubiquitin-proline-LacI, acts as a destabilizing module that permits rapid degradation of appended proteins. Image is Fig S5D, top panels, in J Cell Biol. (2011) 192: 599-614. Images in Fig S5 include CIL # 13880, 13881.

Tem1-CAAX（绿色荧光标记）在野生型细胞中会远离纺锤体极体（spindle pole bodies, SPB）（对应CIL编号13880），但也可定位于SPB（即本图像）。源自Ras2的CAAX基序可将蛋白靶向定位于膜的胞质侧。本实验中，表达eGFP-TEM1-CAAX融合蛋白且携带tem1Δ::GAL-UPL-TEM1基因型的细胞，由着丝粒质粒（CEN plasmid）介导表达，先在2%棉子糖/2%半乳糖培养基中培养，成像前转接至2%葡萄糖培养基中。图像同时包含一张微分干涉差（differential interference contrast, DIC）成像图。tem1Δ::GAL1-UPL-TEM1菌株可实现Tem1的快速条件性降解。UPL全称为泛素-脯氨酸-LacI，作为不稳定模块可快速降解与之融合的靶蛋白。本图像取自《Journal of Cell Biology》2011年第192卷第599-614页的图S5D上半部分。图S5包含的图像对应CIL编号13880与13881。