title: Use of reverse genetics to develop a novel marker porcine reproductive and respiratory syndrome virus.
abstract: Nucleocapsid (N) protein of porcine reproductive and respiratory syndrome virus (PRRSV) is the most abundant viral structural protein with high immunogenicity. Previously, the nonessential sequences for virus infectivity were identified at both N and C terminal ends of N protein. Here, by means of reverse genetics, a marker virus (v7APMa) was generated with a mutant N protein that differs from the wild-type strains (vAPRRS, type 2 PRRSV). v7APMa shows stable inheritance in cell culture and the virologic characteristics of the marker virus in vitro showed that v7APMa replicates well as its parental strain. In the pig model, the v7APMa marker virus induced a similar level of anti-N protein antibodies and robust antibodies against the marker peptide, from 14 days post infection. In addition, a peptide-based ELISA was developed to detect the specific antibodies for the introduced 7APMa peptide. This approach, using a rationally designed marker virus, provides a new potential mutant basis for further development of PRRSV novel vaccines.
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University, No. 1 Weigang Street, Xuanwu District, Nanjing, People's Republic of China.
authors.forename: Tao
authors.initials: T
authors.lastname: Lin
authors.forename: Xiangrui
authors.initials: X
authors.lastname: Li
authors.forename: Huochun
authors.initials: H
authors.lastname: Yao
authors.forename: Zuzhang
authors.initials: Z
authors.lastname: Wei
authors.forename: Feifei
authors.initials: F
authors.lastname: Tan
authors.forename: Runxia
authors.initials: R
authors.lastname: Liu
authors.forename: Lichang
authors.initials: L
authors.lastname: Sun
authors.forename: Rong
authors.initials: R
authors.lastname: Zhang
authors.forename: Wenliang
authors.initials: W
authors.lastname: Li
authors.forename: Jiaqi
authors.initials: J
authors.lastname: Lu
authors.forename: Guangzhi
authors.initials: G
authors.lastname: Tong
authors.forename: Shishan
authors.initials: S
authors.lastname: Yuan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000917:Antibody Formation; D015415:Biomarkers / Q000378:metabolism; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D056724:Immunity, Humoral; D023421:Models, Animal; D008969:Molecular Sequence Data; D016296:Mutagenesis; D019590:Nucleocapsid Proteins / Q000235:genetics* / Q000276:immunology; D016366:Open Reading Frames; D010455:Peptides / Q000235:genetics / Q000276:immunology; D019318:Porcine Reproductive and Respiratory Syndrome / Q000276:immunology / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity / Q000502:physiology; D059386:Reverse Genetics / Q000379:methods*; D013552:Swine; D014162:Transfection; D010948:Viral Plaque Assay; D014779:Virus Replication
chemical_list: D000914:Antibodies, Viral; D015415:Biomarkers; D018076:DNA, Complementary; D019590:Nucleocapsid Proteins; D010455:Peptides