The Borfl protein is encoded by an immediate-early gene of the bovine foamy virus （BFV） and plays a key role in the viral life cycle. Borfl is a DNA binding protein which can transactivate both the long terminal rep...The Borfl protein is encoded by an immediate-early gene of the bovine foamy virus （BFV） and plays a key role in the viral life cycle. Borfl 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 Borfl 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-Borfl serum with a high titer based on ELISA results. Western blot analysis showed that the antiserum could specifically recognize the Borfl protein that was expressed in 293T cells. With this specific serum, we revealed the nuclear and cytoplasmic localization of Borfl in HeLa cells that was transfected with Borfl. Moreover, the immuno-fluorescence assay also showed that the localization of Borfl during the infection and transfection of BFV was identical.展开更多
A 6kb rDNA probe comprising the 18S coding plusspacer sequences has been hybridized to the metaphase chromosomes of apple rootstock cultivar MM106 demon-strating the localization of ribosomal gene arrays in the vicini...A 6kb rDNA probe comprising the 18S coding plusspacer sequences has been hybridized to the metaphase chromosomes of apple rootstock cultivar MM106 demon-strating the localization of ribosomal gene arrays in the vicinity of the telomeric regions of the short arms of chro-mosomes 6 and 14. The in situ results using digoxygenin labelling coupled to an alkaline phosphatase immunoassay were confirmed by silver staining for NORs and nucleoli. This study demonstrates the feasibility of molecular cyto-genetic analysis of very small chromosomes (1.0-2.7μm)of apple.展开更多
基金National Natural Science Foundation ofChina (30570072, 30770097)Natural Science Foun-dation of Tianjin (05YFJM- JC01000).
文摘The Borfl protein is encoded by an immediate-early gene of the bovine foamy virus （BFV） and plays a key role in the viral life cycle. Borfl 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 Borfl 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-Borfl serum with a high titer based on ELISA results. Western blot analysis showed that the antiserum could specifically recognize the Borfl protein that was expressed in 293T cells. With this specific serum, we revealed the nuclear and cytoplasmic localization of Borfl in HeLa cells that was transfected with Borfl. Moreover, the immuno-fluorescence assay also showed that the localization of Borfl during the infection and transfection of BFV was identical.
文摘A 6kb rDNA probe comprising the 18S coding plusspacer sequences has been hybridized to the metaphase chromosomes of apple rootstock cultivar MM106 demon-strating the localization of ribosomal gene arrays in the vicinity of the telomeric regions of the short arms of chro-mosomes 6 and 14. The in situ results using digoxygenin labelling coupled to an alkaline phosphatase immunoassay were confirmed by silver staining for NORs and nucleoli. This study demonstrates the feasibility of molecular cyto-genetic analysis of very small chromosomes (1.0-2.7μm)of apple.
