关键词: 犬瘟热病毒T’TB 抗原表位, 犬细小病毒VP2 基因, 杆状病毒表达系统, 表达, 抗体检测

Abstract: The VP2 gene of canine parvovirus (CPV) was amplified by applying the PCR approach. The amplicon was ligated
into the transfer vector pFastBacHTc of Bac-to-Bac baculovirus expression system. The vector carrying VP2 gene was named pFastBacHTc-VP2. The Artificially synthesized T’TB antigen epitope gene of canine distemper virus (CDV) was inserted into the vector pFastBacHTc-VP2 at the upstream position of VP2 to create a vector carrying both VP2 gene and T’ TB gene named pFastBacHTc-T’TB-VP2. pFastBacHTc-T’TB-VP2 was transferred into E. coli DH10Bac that contains baculovirus shuttle vector bacmid to obtain recombinant transfecting plasmid Bacmid-BacHT-T’TB-VP2. Bacmid-BacHTT’TB-VP2 was then used to transfect Sf-9 insect cells to express recombinant protein T’TB-VP2. Western blot analysis indicated that the molecular weight of T’TB-VP2 was 70 ku and could react with CPV and CDV antiserum. Purified T’TBVP2 protein was used to challenge BALB /c mice at the age of 6 -8 weeks without adjuvant. ELISA testing indicated that T’TB-VP2 protein was able to induce high level of antibodies specifically neutralizing CDV and CPV. This study laid a foundation for developing the new sub-unit vaccine of canine distemper and canine parvovirus.

Key words: Antibody detection, Baculovirus expression system, Expression, Parvovirus virus, T’TB antigen epitope ofcanine distemper virus, VP2 gene of canine