Two coral fluorescent proteins of distinct colors for sharp visualization of cell-cycle progression

We cloned and characterized two new coral fluorescent proteins: h2-3 and 1-41. h2-3 formed an obligate dimeric complex and exhibited bright green fluorescence. On the other hand, 1-41 formed a highly multimeric complex and exhibited dim red fluorescence. We engineered 1-41 into AzaleaB5, a practically useful red-emitting fluorescent protein for cellular labeling applications. We fused h2-3 and AzaleaB5 to the ubiquitination domains of human Geminin and Cdt1, respectively, to generate a new color variant of Fucci (Fluorescent Ubiquitination-based Cell-Cycle Indicator): Fucci5. We found Fucci5 provided more reliable nuclear labeling for monitoring cell-cycle progression than the 1st and 2nd generations that used mAG/mKO2 and mVenus/mCherry, respectively.

我们克隆并鉴定了两种新的珊瑚荧光蛋白：h2 - 3和1 - 41。h2 - 3形成一种必需的二聚体复合物，并呈现出亮绿色荧光。另一方面，1 - 41形成一种高度多聚体复合物，并呈现出暗淡的红色荧光。我们将1 - 41改造成AzaleaB5，这是一种在细胞标记应用中非常实用的发红光的荧光蛋白。我们分别将h2 - 3和AzaleaB5与人Geminin和Cdt1的泛素化结构域融合，以生成一种新的Fucci（基于荧光泛素化的细胞周期指示剂）颜色变体：Fucci5。我们发现，与分别使用mAG/mKO2和mVenus/mCherry的第一代和第二代相比，Fucci5为监测细胞周期进程提供了更可靠的核标记。