Mauricio Valerio-Santiago, Fernando Monje-Casas (2011) CIL:13883, Saccharomyces cerevisiae. CIL. Dataset

Constitutive targeting of Tem1 to the spindle pole body (SPB) in metaphase cells (this image) and anaphase cells (CIL#13886). tem1Δ::GAL-UPL-TEM1 cells expressing eGFP-CNM67–TEM1 from a CEN plasmid were grown on 2% raffinose/2% galactose. Image shows localization of Cnm67-Tem1 (eGFP, green) in metaphase after cells were transferred to medium with 2% glucose. Tem1 normally localizes preferentially to the SPB that enters the daughter cell during anaphase (CIL# 13882, 13885). Cnm67 is an integral component of the outer plaque of the SPB and the Cnm67-Tem1 fusion is found to be symmetric from SPB duplication until the end of mitosis. 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 1A, middle panels, in J Cell Biol. (2011) 192: 599-614. Other images in Fig 1 include CIL #13882, 13883, 13884, 13885, 13886, 13887.

在图示中，Tem1基因对中期细胞（此图像）和后期细胞（CIL#13886）的纺锤体极体（SPB）的构成性靶向进行了研究。tem1Δ::GAL-UPL-TEM1细胞表达eGFP-CNM67–TEM1，该表达来自CEN质粒，并在2%麦芽糖/2%半乳糖培养基中培养。图像展示了细胞转移至2%葡萄糖培养基后，中期Cnm67-Tem1（eGFP，绿色）的定位。Tem1通常优先定位于在后期进入子细胞的SPB（CIL# 13882，13885）。Cnm67是SPB外盘的组成成分，且Cnm67-Tem1融合蛋白在SPB复制直至有丝分裂结束的过程中均表现出对称性。通过DAPI（蓝色）对细胞核形态进行了评估。此外，还展示了一幅微分干涉对比（DIC）图像（灰色）。tem1Δ::GAL1-UPL-TEM1菌株允许快速、条件性地耗竭Tem1。UPL代表泛素-脯氨酸-LacI，它作为一种不稳定的模块，允许快速降解附加蛋白。图像为《细胞生物学杂志》（2011年，第192卷，第599-614页）中的图1A中间面板。图1中的其他图像包括CIL #13882，13883，13884，13885，13886，13887。