Development of a Subunit Vaccine for Canine Parvovirus using Baculovirus Surface Display System

• Main Authors: Chih-Hung Tsai, 蔡志鴻
• Other Authors: Hung-Jen Liu
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/32349016945891540797

碩士 === 國立屏東科技大學 === 動物疫苗科技研究所 === 97 === Canine parvovirus enteritis is a highly contagious disease, the infection is characterized by vomiting、diarrhea and even death. According to previous studies among the canine parvovirus (CPV) capsid proteins, VP2 is the major antigenic determinant that induces neutralizing antibody production. In this study, the full-length VP2 gene of CPV is cloned into the new developed pBacSC vector, containing baculovirus transmembrane domain (gp64 TM) gene, baculovirus cytoplasmic domain (gp64 CTD) gene, green florescence protein (GFP) gene and four multiple cloning sites. GFP can be used as a reporter gene. Gp64 TM and gp64 CTD are able to express heterologous genes on the baculovirus envelope. After cloning the VP2 gene of CPV into pBacSC vector, the recombinant plasmid pBacSC-VP2 is transformed into E. coli DH10Bac competent cells to form recombinant bacmid DNA. The recombinant bacmid DNA is transfected into insect cells, forming “recombinant baculoviruses BacSC-VP2”, expressing the VP2 protein on its envelope. Western blot, confocal microscopy, and immunogold microscopy are used to confirm whether VP2 expressing on baculovirus envelope was successful. The recombinant baculoviruses were injected into mice. The mouse serum was collected and analyzed to check the mouse immune response to the recombinant VP2 protein. The results show that the recombinant baculoviruses could induce good immune response in mice. Our results suggest that the recombinant baculovirus BacSC-VP2 may be used as a subunit for canine parvovirus vaccine.