Samal S K, Pastey M K, McPhillips T H, Mohanty S B
College of Veterinary Medicine, University of Maryland, College Park 20742.
Virology. 1993 Mar;193(1):470-3. doi: 10.1006/viro.1993.1148.
The coding region of the gene for the nucleocapsid (N) protein of bovine respiratory syncytial virus has been inserted into the genome of Autographa californica nuclear polyhedrosis virus (AcNPV) using transfer vector pVL 1393. Infection of Spodoptera frugiperda cells with recombinant virus resulted in the synthesis of high levels of N protein. This protein was indistinguishable from the authentic bovine respiratory syncytial virus (BRSV) N protein by SDS-gel electrophoresis and immunoprecipitation with anti-BRSV serum. [35S]methionine-labeled N protein synthesized in S. frugiperda cells was used in a protein-blotting protein overlay assay for its interactions with BRSV-infected cell proteins. The N protein synthesized in S. frugiperda cells was found to specifically interact with the phosphoprotein (P) and M2 protein of BRSV but only with the P protein of human RSV. Our results suggest that the recombinant N protein produced in S. frugiperda cells could be used to determine the domains on N protein required for binding to P and M2 proteins.
利用转移载体pVL 1393，已将牛呼吸道合胞病毒核衣壳（N）蛋白基因的编码区插入到苜蓿银纹夜蛾核型多角体病毒（AcNPV）的基因组中。用重组病毒感染草地贪夜蛾细胞导致高水平N蛋白的合成。通过SDS-凝胶电泳以及用抗牛呼吸道合胞病毒血清进行免疫沉淀，该蛋白与天然牛呼吸道合胞病毒（BRSV）N蛋白无法区分。在草地贪夜蛾细胞中合成的[35S]甲硫氨酸标记的N蛋白用于蛋白质印迹蛋白覆盖分析，以研究其与感染BRSV的细胞蛋白的相互作用。发现草地贪夜蛾细胞中合成的N蛋白与BRSV的磷蛋白（P）和M2蛋白特异性相互作用，但仅与人呼吸道合胞病毒的P蛋白相互作用。我们的结果表明，在草地贪夜蛾细胞中产生的重组N蛋白可用于确定N蛋白上与P和M2蛋白结合所需的结构域。
