Cloning and Expression of SARS-CoV Related Proteins

• Main Authors: Chih-Jung Ke, 柯志嶸
• Other Authors: Kuang-Hui Sun
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/19693100269389696385

碩士 === 國立陽明大學 === 醫學生物技術研究所 === 94 === The global outbreak of severe acute respiratory syndrome (SARS) occurred in the spring of 2003. The etiology agent was then identified as a novel coronavirus termed SARS-CoV. The spike (S) protein of SARS-CoV is a strong antigen and has been considered to be responsible for virus entry. To determine whether this protein is applicable in vaccine development, clinical treatment, and diagnosis, SARS-CoV spike related proteins were cloned and expressed in this study. The full-length of SI (amino acids 1-460) and SII (amino acids 417-846) were successfully cloned into the Pichia pastoris expression vector. Moreover, HR1 (amino acids 903-952) and HR2 (amino acids 1148-1187) were successfully cloned, expressed and purified in the Escherichia coli expression system. To confirm the functions of HR1 and HR2 recombinant proteins, these proteins were individually added into a cell-to-cell fusion model. The HR2 recombinant protein may reduce the fusion of 293T cells transfected with S protein and those with ACE2 in a dose-dependent manner (2-50 µM) up to 56%. However, this character was not found in the HR1 recombinant protein. Although SARS-CoV spike related proteins were successfully cloned and expressed and the functions of recombinant proteins were also confirmed, further investigations are required to characterize these proteins.