The C-Terminal Portion of the Nucleocapsid Protein Demonstrates SARS-CoV Antigenicity

Liu, Guozhen; Hu, Shaohui; Hu, Yongwu; Chen, Peng; Yin, Jianning; Wen, Jie; Wang, Jingqiang; Lin, Liang; Liu, Jinxiu; You, Bo; Yin, Ye; Li, Shuting; Wang, Hao; Ren, Yan; Ji, Jia; Zhao, Xiaoqian; Sun, Yongqiao; Zhang, Xiaowei; Fang, Jianqiu; Wang, Jian; Liu, Siqi; Yu, Jun; Zhu, Heng; Yang, Huanming

In order to develop clinical diagnostic tools for rapid detection of SARS-CoV (severe acute respiratory syndrome-associated coronavirus) and to identify candidate proteins for vaccine development, the C-terminal portion of the nucleocapsid (NC) gene was amplified using RT-PCR from the SARS-CoV genome, cloned into a yeast expression vector (pEGH), and expressed as a glutathione S-transferase (GST) and Hisx6 double-tagged fusion protein under the control of an inducible promoter. Western analysis on the purified protein confirmed the expression and purification of the NC fusion proteins from yeast. To determine its antigenicity, the fusion protein was challenged with serum samples from SARS patients and normal controls. The NC fusion protein demonstrated high antigenicity with high specificity, and therefore, it should have great potential in designing clinical diagnostic tools and provide useful information for vaccine development.