[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.
Key Laboratory of Veterinary Parasitology of Gansu Province, State Key Laboratory of Veterinary Etiological Biology, CAAS, Lanzhou 730046, Gansu, China.
Yi-Xia
YX
Chen
Xue-Peng
XP
Cai
Zhi-Zhong
ZZ
Jing
Jun-Tao
JT
Ding
Ying
Y
Wang
Xue-Lian
XL
Meng
Yan
Y
Zhang
Wan-Zhong
WZ
Jia
Jun
J
Qiao
Hong-Bin
HB
Yan
Yong-Xiang
YX
Fang
Guo-Hua
GH
Chen
Xue-Nong
XN
Luo
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