Subcellular localization analysis of bovine foamy virus Borf1 protein
Subcellular localization analysis of bovine foamy virus Borf1 protein
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DOI：
10.1007/s12250-008-2893-3
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发表时间：
2008-03
影响因子：
5.5
通讯作者：
Juan Tan;Kai Wu;Rui Chang;Qi-min Chen;Y. Geng;W. Qiao
中科院分区：
文献类型：
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作者：
Juan Tan;Kai Wu;Rui Chang;Qi-min Chen;Y. Geng;W. Qiao
Borf1蛋白由牛泡沫病毒(BFV)的即刻早期基因编码，在病毒生命周期中发挥关键作用。Borf1是一种DNA结合蛋白，通过与反式激活反应元件(Tre)特异性结合，可以反式激活BFV的长端重复序列(LTR)和内部启动子(IP)。为了分析Borf1基因在BFV生活史中的亚细胞定位，将该基因克隆到原核表达载体中，并以可溶性形式表达。纯化和免疫后，根据ELISA法获得高滴度的小鼠抗Borf1血清。Western印迹分析表明，该抗血清能特异性识别293T细胞中表达的Borf1蛋白。用此特异性血清，我们揭示了Borf1在转染Borf1的HeLa细胞中的核和胞质定位。此外，免疫荧光分析还表明，Borf1在BFV感染和转染过程中的定位是一致的。
The Borf1 protein is encoded by an immediate-early gene of the bovine foamy virus (BFV) and plays a key role in the viral life cycle. Borf1 is a DNA binding protein which can transactivate both the long terminal repeat (LTR) and the internal promoter (IP) of BFV by specifically binding to the transactivation responsive element (TRE). To analyze the subcellular localization of Borf1 during the BFV life cycle, this gene was cloned into a prokaryotic expression vector and expressed in a soluble form. After the purification and immunization, we raised the mouse anti-Borf1 serum with a high titer based on ELISA results. Western blot analysis showed that the antiserum could specifically recognize the Borf1 protein that was expressed in 293T cells. With this specific serum, we revealed the nuclear and cytoplasmic localization of Borf1 in HeLa cells that was transfected with Borf1. Moreover, the immuno-fluorescence assay also showed that the localization of Borf1 during the infection and transfection of BFV was identical.