CIL:13885

Tem1 normally localizes preferentially to the spindle pole body (SPB) that enters the daughter cell during anaphase (this image and CIL# 13882). This image shows SPB localization of eGFP-Tem1 (eGFP, green) in anaphase cells. tem1Δ::GAL-UPL-TEM1 cells expressing eGFP-TEM1 from a CEN plasmid were grown on 2% raffinose/2% galactose and transferred to 2% glucose medium prior to image capture. Nuclear morphology was assessed by DAPI (blue). A differential interference contrast (DIC) image is also shown (gray). 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 1B, top panels, in J Cell Biol. (2011) 192: 599-614. Other images in Fig 1 include CIL #13882, 13883, 13884, 13885, 13886, 13887.Tem1通常优先定位于有丝分裂后期进入子细胞的纺锤体极体（spindle pole body，SPB），本图像与CIL#13882均为该类定位的示例。本图像展示了有丝分裂后期细胞中eGFP标记的Tem1（增强绿色荧光蛋白，enhanced green fluorescent protein，eGFP，绿色荧光）在纺锤体极体上的定位情况。携带着丝粒质粒（centromere plasmid，CEN plasmid）表达eGFP-TEM1的tem1Δ::GAL-UPL-TEM1菌株，先在含2%棉子糖与2%半乳糖的培养基中培养，随后转移至2%葡萄糖培养基中，再进行图像采集。采用4',6-二脒基-2-苯基吲哚（4',6-diamidino-2-phenylindole，DAPI，蓝色荧光）标记以评估细胞核形态。同时还展示了一幅微分干涉差显微镜（differential interference contrast，DIC）灰度图像。tem1Δ::GAL1-UPL-TEM1菌株可实现Tem1的快速条件性降解。UPL即泛素-脯氨酸-LacI（ubiquitin-proline-LacI），作为一种去稳定结构域，可介导融合蛋白的快速降解。本图像出自《J Cell Biol》2011年第192卷第599至614页的图1B上方面板。图1中的其他图像还包括CIL#13882、13883、13884、13885、13886及13887。