A fluorescent p53GFP fusion protein facilitates its detection in mammalian cells while retaining the properties of wild-type p53.
A fluorescent p53GFP fusion protein facilitates its detection in mammalian cells while retaining the properties of wild-type p53.
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荧光 p53GFP 融合蛋白有助于其在哺乳动物细胞中的检测，同时保留野生型 p53 的特性。
DOI：
10.1038/sj.onc.1201406
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Haas,M
中科院分区：
文献类型：
--
作者：
Norris,PS;Haas,M
肿瘤进展通常以关键细胞周期控制基因的累积丢失和伴随的基因组稳定性丢失为特征。进展性肿瘤通常对常规疗法具有抗性。关键生长调节基因（如p53基因）的基因转移是治疗晚期肿瘤的一种潜在方法。为此，我们已经产生了高滴度的逆转录病毒，基于pCL载体系统，其编码由人野生型p53和绿色荧光蛋白（wtp53 GFP）组成的嵌合蛋白。荧光wtp53GFP蛋白和野生型p53蛋白同样被几种单克隆p53特异性抗体识别，在反式激活p53反应元件以及抑制肿瘤细胞生长方面具有相似的半衰期和功能。此外，由于其荧光性质，wtp53GFP有利于直接鉴定表达p53融合蛋白的细胞。将pCL逆转录病毒生产系统的特征与高度可见的绿色荧光蛋白相结合，为将p53递送到细胞中以及随后在体外和体内检测蛋白质提供了有力的工具。
Tumor progression is often characterized by the cumulative loss of crucial cell cycle control genes and the concomitant loss of genome stability. Progressed tumors are often resistant to conventional therapies. Gene-transfer of key growth-regulatory genes, such as the p53 gene, is one potential approach to treating advanced tumors. To this end, we have produced high-titer retroviruses, based on the pCL vector system, which encode a chimeric protein consisting of human wild-type p53 and the green fluorescent protein (wtp53GFP). The fluorescent wtp53GFP protein and the wild-type p53 protein are recognized equally by several monoclonal p53-specific antibodies, have similar half-lives and function comparably in transactivating a p53-responsive element as well as in suppressing the growth of tumor cells. Additionally, due to its fluorescent nature, wtp53GFP facilitates the direct identification of cells expressing the p53 fusion protein. Combining the features of the pCL retroviral production system with the highly visible green fluorescent protein provides a potent tool for the delivery of p53 into cells and the subsequent detection of the protein, both in vitro and in vivo.