Yang P, Flügel R M, Li W, Löchelt M
Institute of Virology, College of Life Science, Wuhan University, Hubei, PR China.
Virus Genes. 2000 Oct;21(3):209-13. doi: 10.1023/a:1008143615032.
Recombinant plasmids that express human foamy virus (HFV) Bel 1 transactivator and human estrogen receptor (ER) fusion proteins were constructed. The HFV bel 1 gene was inserted up- and downstream of the ER gene. Recombinant Bel 1-ER and ER-Bel 1 fusion proteins were expressed in eukaryotic cells. In the absence of estrogen, the ER moiety of the fusion proteins suppressed Bel 1-mediated transactivation as measured in CAT reporter gene-based transactivation assays. However, transactivation of the HFV LTR and the HFV internal promoter by Bel 1-ER and ER-Bel 1 fusion proteins was recovered in the presence of estrogen. Thus, the transactivation function of the Bel 1 moiety of the chimeric Bel 1-ER fusion proteins can be efficiently, specifically, and intentionally activated and inactivated by simply adding the low-molecular weight effector estrogen.
构建了表达人泡沫病毒(HFV)Bel 1反式激活因子与人雌激素受体(ER)融合蛋白的重组质粒。将HFV bel 1基因插入ER基因的上下游。重组Bel 1-ER和ER-Bel 1融合蛋白在真核细胞中表达。在缺乏雌激素的情况下,基于CAT报告基因的反式激活分析显示,融合蛋白的ER部分抑制了Bel 1介导的反式激活。然而,在存在雌激素的情况下,Bel 1-ER和ER-Bel 1融合蛋白对HFV LTR和HFV内部启动子的反式激活得以恢复。因此,通过简单添加低分子量效应物雌激素,嵌合Bel 1-ER融合蛋白的Bel 1部分的反式激活功能可以被有效、特异性且有意地激活和失活。