口蹄疫病毒多基因及猪α干扰素共表达载体的构建及其免疫豚鼠效果研究

doi:10.16431/j.cnki.1671-7236.2017.05.028

Abstract

Abstract:

In order to explore the feasibility of entering clinical trials of multi-gene of foot-and-mouth disease virus (FMDV) and porcine interferon-α(IFN-α) co-expression plasmids, porcine IFN-α and the multi-gene of FMDV that included full length of P12A3C,and part of 2B (P12X3C) were amplified by PCR and cloned into the eukaryotic expressing plasmid pBudCE4.1 to construct the recombinant vector. After identification by restriction enzyme digestion, this vector was transfected into the BHK-21 cell and its expression were observed by Western blotting and indirect fluoroscopy assay.The serum antibodies, neutralizing antibody titers and T-lymphocyte proliferation were detected in guinea pigs after immunization of the recombinant vector. The results showed that the recombinant plasmid pBudCE4.1-P12X3C-IFN-α was successfully constructed after detecting by enzyme digestion and sequencing. Recombinant plasmid was effectively expressed in BHK-21 cells after detecting by Western blotting and indirect fluoroscopy assay. Result of ELISA indicated that anti-FMDV IgG antibody level of pBudCE4.1-P12X3C-IFN-α group was higher compared with pBudCE4.1-P12X3C group.Similar results for serum neutralization titers were obtained. MTT assay showed that proliferation of lymphocyte was 15% in pBudCE4.1-P12X3C-IFN-α group and 11% in case of pBudCE4.1-P12X3C group. After challenged with homologous virus, protection rate of conventional vaccine and recombinant plasmid pBudCE4.1-P12X3C-IFN-α group were 100%, while the recombinant plasmid pBudCE4.1-P12X3C group was 80%.The present study showed the recombinant plasmid pBudCE4.1-P12X3C-IFN-α was successfully constructed, and could markedly improve the efficacy of DNA vaccine against FMDV.

Key words: porcine IFN-α; FMDV; eukaryotic expression vector; guinea pig

CLC Number:

S852.65

ZHENG Hua-bin, ZHANG Zhong-wang, LV Jian-liang, PAN Li, ZHANG Yong-guang. Construction of Co-expression Vector Containing Multi-gene of FMDV and Porcine IFN-α and Its Immunogenicity Effect in Guinea Pig[J]. , 2017, 44(5): 1452-1461.

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Construction of Co-expression Vector Containing Multi-gene of FMDV and Porcine IFN-α and Its Immunogenicity Effect in Guinea Pig

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