Construction, expression and immunogenicity of eukaryotic vectors based on goat pox virus P32 gene.
- Author:
Yi-Xia CHEN
1
;
Xue-Peng CAI
;
Zhi-Zhong JING
;
Jun-Tao DING
;
Ying WANG
;
Xue-Lian MENG
;
Yan ZHANG
;
Wan-Zhong JIA
;
Jun QIAO
;
Hong-Bin YAN
;
Yong-Xiang FANG
;
Guo-Hua CHEN
;
Xue-Nong LUO
Author Information
1. Key Laboratory of Veterinary Parasitology of Gansu Province, State Key Laboratory of Veterinary Etiological Biology, CAAS, Lanzhou 730046, Gansu, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Viral;
blood;
Capripoxvirus;
genetics;
immunology;
Cell Line;
CpG Islands;
Cricetinae;
Female;
Male;
Mice;
Mice, Inbred BALB C;
Recombinant Proteins;
immunology;
T-Lymphocyte Subsets;
immunology;
Vaccines, Synthetic;
immunology;
Viral Envelope Proteins;
immunology;
Viral Vaccines;
immunology
- From:
Chinese Journal of Virology
2008;24(2):133-137
- CountryChina
- Language:Chinese
-
Abstract:
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.
