共表达牛病毒性腹泻病毒E0基因和牛传染性鼻气管炎病毒gD基因的重组质粒DNA构建及其对小鼠的免疫效应

doi:10.11843/j.issn.0366-6964.2021.016

Abstract

Abstract: This study aimed to construct a recombinant plasmid that can simultaneously express the dominant antigen genes of bovine viral diarrhea virus (BVDV) and infectious bovine rhinotracheitis virus (IBRV), and study its immune effect, so as to provide reliable data and theoretical support for the development of BVD-IBR bivalent DNA vaccine. In our experiment, PCR was used to amplify the target genes, E0 and gD, which were sequentially connected to the eukaryotic expression vector pVAX1-IRES to construct the pVAX1-E0-IRES-gD recombinant eukaryotic expression vector. The recombinant plasmid pVAX1-E0-IRES-gD was transfected into 293T cells and the expression of the target genes in foreign cells were detected by indirect immunofluorescence assay. Following, immunization test with the pVAX1-E0-IRES-gD, pVAX1-IRES and BVD-IBR combined inactivated vaccine was inoculated with SPF mice at different doses by intramuscular injection, subsequently, immune effects have been evaluated through antibody level and cytokine detection and splenic lymphocyte proliferation assay. The results showed that the pVAX1-E0-IRES-gD recombinant plasmid was successfully constructed and both antigen genes obtained a high level of expression in 293T cells. In terms of antibody and cytokine levels and the proliferation of spleen lymphocytes, the recombinant plasmid group was significantly higher than that of the empty vector and the negative control group, and the differences were extremely significant, the high-dose immunization group was better than the medium and low dose groups. Also, the high-dose recombinant plasmid immunization group was not significantly different from the BVD-IBR inactivated vaccine group. The results showed that the co-expression recombinant plasmid which contains the BVDV E0 gene and IBRV gD gene was constructed successfully. The expression test in vitro proved that the target protein has good reactionogenicity, and animal immunity tests proved that it can stimulate the body to produce a good immune response.

Key words: bovine viral diarrhea virus, E0, infectious bovine rhinotracheities virus, gD, recombinant plasmid, co-expression, immune effect

CLC Number:

S858.235.3

MA Xiaojing, LI Mengying, ZHANG Huaiyu, SONG Abei, XU Lihua, WU Xinhua, ZHANG Qiaoe, LI Jidong. Construction of a Recombinant Plasmid Co-expressing Bovine Viral Diarrhea Virus E0 Gene and Infectious Bovine Rhinotracheitis Virus gD Gene and Consider Its Immune Effect on Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(1): 154-165.

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Construction of a Recombinant Plasmid Co-expressing Bovine Viral Diarrhea Virus E0 Gene and Infectious Bovine Rhinotracheitis Virus gD Gene and Consider Its Immune Effect on Mice

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