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Attenuated porcine-derived type 2 bovine viral diarrhea virus as vector stably expressing viral gene

文献类型： 外文期刊

作者： Tao, Jie 1 ; Li, Benqiang 1 ; Shi, Ying 1 ; Chen, Jinghua 1 ; Zhu, Guoqiang 3 ; Shen, Xiaohui 1 ; Liu, Huili 1 ;

作者机构： 1.Shanghai Acad Agr Sci, Inst Anim Sci & Vet Med, BeiDi Rd 2901, Shanghai, Peoples R China

2.Shanghai Key Lab Agr Genet Breeding, Shanghai 201106, Peoples R China

3.Yangzhou Univ, Coll Vet Med, Yangzhou 225009, Jiangsu, Peoples R China

4.Yangzhou Univ, Jiangsu Coinnovat Ctr Prevent & Control Important, Yangzhou 225009, Jiangsu, Peoples R China

5.Shanghai Engn Res Ctr Pig Breeding, Shanghai 201302, Peoples R China

关键词： BVDV; Pig; PEDV; Chimeric virus; Vaccine carrier; Immunogenicity

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN： 0166-0934

年卷期： 2020 年 279 卷

页码：

收录情况： SCI

摘要： Infectious bovine viral diarrhea virus (BVDV) cDNA clones have been used for the expression of classical swine fever virus (CSFV) genes for immune prevention and control. However, can it be used for the expression of an allogenetic fragment? To answer this question, a BVDV chimeric virus expressing the spike (S) antigen fragment of porcine epidemic diarrhea virus (PEDV) was constructed. Antigen S499-602 was inserted into pig-derived BVDV-2 infectious cDNA clone pASH28 in tandem by overlapping PCR, located between the seventh and eighth amino acids at the N-terminus of the capsid (C) protein of BVDV. Indirect immunofluorescence assay confirmed that the chimeric virus vASH-dS312 containing double S499-602 sequences was successfully assembled, which could react with the monoclonal antibody (MAb) against BVDV E2 and PEDV S proteins. Further western blot analysis confirmed that the exogenous S499-602 double protein could be stably expressed. Next, the chimeric virus vASH-dS312 was administered to BALB/C mice either orally or by intramuscular injection. The immunized mice were healthy and showed no signs of toxicity. IgG against BVDV and PEDV antibodies could be detected in the mice administered vASH-dS312 by intramuscular injection, which had neutralization activity against BVDV and PEDV. Thus, this study reported a new insertion site in the BVDV infectious cDNA clone that could successfully express an allogenetic antigen.

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