Cyclin B1 destruction box-mediated protein instability: the enhanced sensitivity of fluorescent-protein-based reporter gene system

细胞周期蛋白B1破坏盒介导的蛋白质不稳定性:基于荧光蛋白的报告基因系统的增强敏感性

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作者:Chao-Hsun Yang, Wan-Ting Kuo, Yun-Ting Chuang, Cheng-Yu Chen, Chih-Chien Lin

Abstract

The periodic expression and destruction of several cyclins are the most important steps for the exact regulation of cell cycle. Cyclins are degraded by the ubiquitin-proteasome system during cell cycle. Besides, a short sequence near the N-terminal of cyclin B called the destruction box (D-box; CDB) is also required. Fluorescent-protein-based reporter gene system is insensitive to analysis because of the overly stable fluorescent proteins. Therefore, in this study, we use human CDB fused with both enhanced green fluorescent protein (EGFP) at C-terminus and red fluorescent protein (RFP, DsRed) at N-terminus in the transfected human melanoma cells to examine the effects of CDB on different fluorescent proteins. Our results indicated that CDB-fused fluorescent protein can be used to examine the slight gene regulations in the reporter gene system and have the potential to be the system for screening of functional compounds in the future.

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