College of Life Science and Technology, Beijing University of Chemical Technology, 100029, Beijing, China,
Cytotechnology. 2010 Apr;62(2):133-41. doi: 10.1007/s10616-010-9269-x. Epub 2010 Apr 21.
Virus-templated fabrication of compound structures can be made through incorporating the specifically inorganic-binding peptide into the viral scaffold, widely used is phage display system. Compared to prokaryotic phages, insect cell-based baculovirus has some strengths such as the adaptability to the proteins' posttranslational modification and non-replication in mammalian cells. As an attempt to explore the baculovirus-mediated bioconjugates, we show in this study that a genetically engineered baculovirus, with a hexahistidine (His(6)) tagged ZnO binding peptide fused to the N-terminus of the viral capsid protein vp39 of AcNPV, was constructed. It maintains both the viral infectivity and the fusion protein's activity. The presence of the fusion protein on the baculovirus particle was demonstrated by western blot analysis of purified budded virus. Its display on the virus capsid was revealed by virus fractionation analysis. The binding of nanosized ZnO powders to the virus capsid was visualized by transmission electron microscopy (TEM). This is the first report of the display of the inorganic-binding peptide on the capsid of eukaryotic baculovirus. Aimed at the nanomaterials' application in the biological field, this research could find useful in the biotracking of the baculovirus transduction process and the preparation of novel functional nanodevices.
病毒模板制备化合物结构可以通过将特定的无机结合肽整合到病毒支架中实现,其中广泛使用的是噬菌体展示系统。与原核噬菌体相比,基于昆虫细胞的杆状病毒具有一些优势,例如能够适应蛋白质的翻译后修饰,并且在哺乳动物细胞中不能复制。作为探索杆状病毒介导的生物缀合物的尝试,本研究构建了一种遗传工程杆状病毒,该病毒将六组氨酸(His(6))标记的 ZnO 结合肽融合到杆状病毒 AcNPV 的衣壳蛋白 vp39 的 N 端。它保持了病毒的感染力和融合蛋白的活性。通过对纯化的出芽病毒进行 Western blot 分析证明了融合蛋白存在于杆状病毒颗粒上。通过病毒分级分析揭示了其在病毒衣壳上的展示。通过透射电子显微镜(TEM)观察到纳米 ZnO 粉末与病毒衣壳的结合。这是首次报道真核杆状病毒衣壳上展示无机结合肽。针对纳米材料在生物领域的应用,本研究可用于杆状病毒转导过程的生物追踪和新型功能纳米器件的制备。