[Construction, expression and immunogenicity of eukaryotic vectors based on goat pox virus P32 gene].
The full-length P32 gene and the truncated P32 gene (MP-32) were amplified from the recombinant plasmid pMD-P32 by polymerase chain reaction (PCR) and cloned into pcDNA3. 1(+) and pcDNA3.1-CpG respectively. The recombinant plasmids (pcDNA3.1-P32, pcDNA3.1-CpG-P32 and pcDNA3. 1-CpG-MP32) were transfected into BHK-21 cells by using lipofectin. The expressed P32 protein was confirmed by indirect immunofluorescence assay (IFA). The BALB/c mice were immunized with these recombinant plasmids by intramuscular injection. The specific antibodies aginst CPV were detected by ELISA kit weekly. The murine splenic T lymphocyte subgroups CD4+ and CD8+ were detected by flow cytometry. Results showed that the P32 protein was expressed successfully in vitro. After 2 weeks post im munization, the specific IgG antibodies against CPV were detected in the vaccinated mice. The percentage of CD4+ /CD8+ T cells was significantly higher than that of the control. In conclusion, these constructed eukaryotic vectors could induce humoral and celluar immune responses in mice.
Chen
Yi-Xia
YX
Key Laboratory of Veterinary Parasitology of Gansu Province, State Key Laboratory of Veterinary Etiological Biology, CAAS, Lanzhou 730046, Gansu, China.
Cai
Xue-Peng
XP
Jing
Zhi-Zhong
ZZ
Ding
Jun-Tao
JT
Wang
Ying
Y
Meng
Xue-Lian
XL
Zhang
Yan
Y
Jia
Wan-Zhong
WZ
Qiao
Jun
J
Yan
Hong-Bin
HB
Fang
Yong-Xiang
YX
Chen
Guo-Hua
GH
Luo
Xue-Nong
XN
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D018151:Capripoxvirus / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D018899:CpG Islands; D006224:Cricetinae; D005260:Female; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014765:Viral Vaccines