Azad A A, Jagadish M N, Brown M A, Hudson P J
CSIRO, Division of Protein Chemistry, Parkville, Australia.
Virology. 1987 Nov;161(1):145-52. doi: 10.1016/0042-6822(87)90180-2.
The large genomic segment of infectious bursal disease virus encodes a polyprotein in which the viral polypeptides are present in the following order: N-VP2-VP4-VP3-C. Expression in Escherichia coli of the large segment results in the processing of the polyprotein. The expression product reacts with a virus neutralizing and protective monoclonal antibody that recognizes a conformational epitope on the surface of the virus. Different regions of the large genomic segment were deleted at defined restriction sites and the truncated fragments were ligated to various expression vectors for high-level expression in E. coli. The expressed proteins were probed with three different monoclonal antibodies that recognize epitopes encoded by different regions of the large genomic segment. These deletion mapping studies suggest that VP4 is involved in the processing of the precursor polyprotein, and the conformational epitope recognized by the virus neutralizing monoclonal antibody is present within VP2.
传染性法氏囊病病毒的大基因组片段编码一种多聚蛋白，其中病毒多肽按以下顺序排列：N-VP2-VP4-VP3-C。在大肠杆菌中表达该大片段会导致多聚蛋白的加工。表达产物与一种病毒中和及保护性单克隆抗体发生反应，该抗体识别病毒表面的一个构象表位。在特定的限制酶切位点删除大基因组片段的不同区域，并将截短的片段连接到各种表达载体上，以便在大肠杆菌中进行高水平表达。用三种不同的单克隆抗体对表达的蛋白质进行检测，这些抗体识别大基因组片段不同区域编码的表位。这些缺失定位研究表明，VP4参与前体多聚蛋白的加工，并且病毒中和单克隆抗体识别的构象表位存在于VP2内。
