Yamada S, Shimizu M
Second Research Division, National Institute of Animal Health, Ibaraki, Japan.
Virology. 1994 Jul;202(1):491-5. doi: 10.1006/viro.1994.1368.
IE180, an immediate-early regulatory protein of pseudorabies virus, is required to activate viral early and late genes and is essential for viral growth. We constructed a recombinant baculovirus expressing IE180 in SF21AE insect cells. IE180 expressed by the recombinant migrated on gels as a single band with a molecular weight of 180 kDa and reacted with anti-IE180 peptide serum in Western blotting assay. Immunofluorescence using anti-IE180 peptide antibody demonstrated the accumulation of IE180 in cytoplasm of SF21AE cells infected with the recombinant virus. The IE180 produced in the insect cells was introduced into mammalian Vero cells infected with a PrV mutant with deletion of the IE180 gene by a cell-fusion technique. The growth of the deletion mutant in the fused cells was confirmed by infective assay and immunodetection of the progeny virus. This indicates that the IE180 expressed by the recombinant baculovirus is functional as a transactivator.
IE180是伪狂犬病病毒的一种立即早期调节蛋白，它是激活病毒早期和晚期基因所必需的，对病毒生长至关重要。我们构建了一种在SF21AE昆虫细胞中表达IE180的重组杆状病毒。重组体表达的IE180在凝胶上迁移为一条分子量为180 kDa的单带，并在蛋白质免疫印迹分析中与抗IE180肽血清发生反应。使用抗IE180肽抗体的免疫荧光显示，IE180在感染重组病毒的SF21AE细胞的细胞质中积累。通过细胞融合技术，将昆虫细胞中产生的IE180导入感染了缺失IE180基因的PrV突变体的哺乳动物Vero细胞中。通过感染性测定和子代病毒的免疫检测证实了缺失突变体在融合细胞中的生长。这表明重组杆状病毒表达的IE180作为反式激活因子具有功能。
