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    • 6. 发明申请
    • N-linked glycosylation alteration in e1 glycoprotein of classical swine fever virus and novel classical swine fever virus vaccine
    • 的糖基化N IN CONNECTION糖蛋白E1 VIRUS猪瘟CLASSIC蚀变和疫苗对病毒
    • WO2009091720A3
    • 2009-10-08
    • PCT/US2009030824
    • 2009-01-13
    • US AGRICULTUREBORCA MANUEL VRISATTI GUILLERMO R
    • BORCA MANUEL VRISATTI GUILLERMO R
    • C12N15/33
    • C07H21/04A61K39/12A61K2039/5254A61K2039/552C12N7/00C12N2770/24334C12N2770/24362G01N33/56983G01N2333/183
    • E1, along with Ems and E2 is one of the three envelope glycoproteins of Classical Swine Fever Virus (CSFV). Our previous studies indicated that glycosylation status of either E2 or Erns strongly influence viral virulence in swine. Here, we have investigated the role of E1 glycosylation of highly virulent CSFV strain Brescia during infection in the natural host. The three putative glycosylation sites in E1 were modified by site directed mutagenesis of a CSFV Brescia infectious clone (BICv). A panel of virus mutants was obtained and used to investigate whether the removal of putative glycosylation sites in the E1 glycoprotein would affect viral virulence/pathogenesis in swine. We observed that rescue of viable virus was completely impaired by removal of all three putative glycosylation sites in E1. Single mutations of each of the E1 glycosylation sites showed that CSFV amino acid N594 (E1.N3 virus), as well the combined mutation of N500 and N513 (E1.N1N2 virus) resulted in BICv attenuation. Infection of either E1. N1N2 or E1.N3 viruses were able to efficiently protected swine from challenge with virulent BICv at 3 and 28 days post-infection. These results, along with those demonstrating the role of glycosylation of Erps and E2, suggest that manipulation of the pattern of glycosylation could be a useful tool for development of CSF live-attenuated vaccines.
    • 根据本发明,E1,和E2用EMS,猪瘟(CSFV)的病毒的三个包膜糖蛋白中的一个。 我们以往的研究表明,糖基化状态E2或Erns的强烈影响猪的病毒毒力。 本发明是对病毒的高毒力株布雷西亚的E1糖基化的天然宿主的感染过程中的作用的研究结果。 在E1的三个假定基化位点被CSFV布雷西亚(BICv)的感染性克隆的定点诱变来修饰。 得到的一组突变体病毒的并用于确定在E1糖蛋白中除去推定的糖基化位点的改变是否在猪的毒力/病毒致病。 据观察,在可行的拯救病毒完全被去除的E1三个假定基化位点改变。 每个E1糖基化位N594的单突变表明CSFV氨基酸(E1.N3病毒),和N500和N513(E1.N1N2病毒)的组合突变导致衰减BICv。 感染病毒或E1.N1N2 E1.N3,有效防止有毒BICv,3和感染28天后的猪。 这些结果,与糖基化的Erns和E2的作用结果相结合表明,糖基化模式的操作是在减弱的猪瘟活疫苗发展的有用工具。