Thumbnail Image

Theses

Browse

20221
Reset filters
Subject: Animal Biotechnology × Author: BORAH, DEBARUN × Type: Thesis ×
Reset filters

Search Results

Now showing 1 - 1 of 1
  • Thumbnail Image
    ThesisItemOpen Access
    EXPRESSION OF CAP PROTEIN OF PORCINE CIRCOVIRUS TYPE 2 (PCV2) AND EVALUATION OF ITS IMMUNOGENICITY IN MICE
    (College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati, 2022-03) BORAH, DEBARUN; Hazarika, Girin
    The present study was conducted with an aim to express the Cap protein of Porcine Circovirus Type 2 (PCV2) in a prokaryotic host and to assess its immunogenicity in mice. The recombinant pET32a/CT-His plasmid containing a codon-optimised PCV2 cap gene was transformed into E. coli BL21(DE3) cells for expression of the Cap protein after 8 hours of IPTG induction at 37°C. Subsequent SDS-PAGE analysis revealed an intense band of 46 kDa. Following successful purification by affinity chromatography using Ni-NTA column under denaturing conditions, the recombinant protein was refolded by dialysis. In western blot analysis of the purified protein, a 46.0 kDa-sized intense band was detected in the PVDF membrane. In four groups of Swiss albino mice consisting of six mice in each group, the immunological response to the purified recombinant Cap protein was assessed and compared to that of a commercial vaccine. Group 1 served as the control group receiving PBS, while Group 2 received a commercial vaccine, Group 3 received the recombinant Cap protein, and Group 4 received the Cap protein combined with Freund's adjuvant. On the 14th day post-primary immunization, each group of mice received a booster dose of the corresponding inoculum. Serum was collected from all the experimental animals on 0, 7th, 14th, 21st, 28th, 35th, and 42nd days after immunization. As detected by indirect ELISA, the mean antibody level against PCV2 was significantly higher in Group 4 getting Cap protein with Freund's adjuvant, than in Group 2 receiving the commercial vaccine, and Group 3 receiving the Cap protein alone. The recombinant Cap protein was found to be immunogenic in mice and was capable of inducing a specific IgG response, as detected by indirect ELISA. Based on the results of the present study, it may be concluded that the recombinant Cap protein expressed in E. coli is a promising immunogen and may be further explored as a vaccine candidate against PCV2. Use of the recombinant protein as a potential diagnostic antigen may also be explored in future.