TY  - JOUR
T1  - Expression of CSFV E2 Protein in Eukaryotic Cells and Characterization of 
  Immunity in Animals
AU - Zhang, Yan-Ming AU - Liu, Jian-Ling AU - Guo, Kang-Kang AU - Su, Zheng-Yuan AU - Xu, Xin-Gang AU - Li, He-Lin AU - Ning, Peng-Bo AU - Hong, Hai-Xia AU - Yang, Xiao-Yun 
JO  - Journal of Animal and Veterinary Advances
VL  - 11
IS  - 18
SP  - 3312
EP  - 3317
PY  - 2012
DA  - 2001/08/19
SN  - 1680-5593
DO  - javaa.2012.3312.3317
UR  - https://makhillpublications.co/view-article.php?doi=javaa.2012.3312.3317
KW  - Classical Swine fever virus
KW  -E2 protein
KW  -retroviral vector
KW  -eukaryotic expression
KW  -immune protection experiment
AB  - The viral envelope glycoprotein E2 is major target for eliciting protective antibodies against CSFV in infected animals. To express CSFV E2 protein on eukaryotic cells, the recombinant retroviral vector pBabe-puro-E2 was constructed by inserting full-length CSFV Shimen strain E2 region into pBabe-puro. Both of the recombinant retroviral vector and pVSVg plasmid were transfected eukaryotic 293GP cells using method of calcium phosphate coprecipitation in where thus the recombinant pseudovirus were packaged and propagated. The PK-15 cells were infected with recombinant pseudovirus. The recombinant PK-15 cells were screened by spytomycin resistance and the expression of E2 protein was detected by Flow Cytometer (FCM). The activity of recombinant E2 protein to induce immune responses was evaluated in Balb/c mice and unvaccinated pigs. The results showed that CSFV E2 protein was successfully expressed on PK-15 cells membrane. Anti-E2 antibody induced in experimental animals was detected by Enzyme Linked Immunosorbent Assay (ELISA). Moreover, the virus challenge indicated that the immunized pigs generated effective protection against virulent CSFV. These results indicated that a retroviral-based E2-vaccine is able to induce high level of specific antibodies and exhibits similar protective capability with that induced by the C-strain.
ER  -