title: An attenuated EMCV-HB10 strain acts as a live viral vector delivering a foreign gene.
abstract: We successfully constructed a full-length cDNA infectious clone of the encephalomyocarditis virus (EMCV) HB10 strain and obtained a partially attenuated rEMCV-C9 virus with a shorter poly(C) tract. Our results showed that the length of the EMCV-HB10 poly(C) tract was related to the pathogenicity of the EMCV-HB10 strain in vivo. Using pEMCV-C9 as the backbone, we constructed the novel viral vector pC9-MCS-∆2A by inserting a cDNA fragment containing a 127 amino acid deletion in the 2A protein, a primary cleavage cassette, a FLAG tag and a multiple cloning site (MCS) at the junction of VP1 and ∆2A. Additionally, the enhanced green fluorescent protein (egfp) gene was cloned into the MCS of pC9-MCS-∆2A to test its capacity to express foreign proteins. Insertion of the egfp gene did not affect viral replication, and a decrease in EGFP expression was observed within five serial passages. Furthermore, we found that rC9-EGFP-∆2A was avirulent in vivo, induced neutralizing antibody production and conferred protective immune responses against lethal challenge with EMCV in mice. Taken together, our results demonstrated that we had constructed an attenuated live vector based on an EMCV-HB10 strain with two modified critical virulence factors (the poly(C) tract and 2A protein) that could be used as a candidate live vaccine and a potential live viral vector for foreign antigen delivery.
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, PR China.
authors.forename: Huibin
authors.initials: H
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, PR China.
authors.forename: Li
authors.initials: L
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, PR China.
authors.forename: Yuanfeng
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, PR China.
authors.forename: Liang
authors.initials: L
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, PR China.
authors.forename: Shengnan
authors.initials: S
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, PR China.
authors.forename: Jiangnan
authors.initials: J
authors.lastname: Li
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, PR China.
authors.forename: Xuehui
authors.initials: X
authors.lastname: Cai
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, PR China.
authors.forename: Shangjin
authors.initials: S
authors.lastname: Cui
authors.affiliation: State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, PR China.
authors.forename: Changjiang
authors.initials: C
authors.lastname: Weng
mesh_terms: D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004337:Drug Carriers; D004686:Encephalomyocarditis virus / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D042822:Genomic Instability; D051379:Mice; D008967:Molecular Biology / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012692:Serial Passage; D013678:Technology, Pharmaceutical / Q000379:methods; D014613:Vaccines, Attenuated / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014779:Virus Replication
chemical_list: D018076:DNA, Complementary; D004279:DNA, Viral; D004337:Drug Carriers; D011994:Recombinant Proteins; D014613:Vaccines, Attenuated; D014614:Vaccines, Synthetic