Immunogenicity of the capsid precursor and a nine-amino-acid site-directed mutant of the 3C protease of foot-and-mouth disease virus coexpressed by a recombinant goatpox virus

文献类型： 外文期刊

第一作者： Ma, Wenge

作者： Ma, Wenge;Wei, Jie;Wei, Yurong;Guo, Huiling;Jin, Yinghong;Xue, Ying;Wang, Yan;Yi, Zhong;Liu, Liya;Huang, Jiong;Wang, Lijian

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期刊名称：ARCHIVES OF VIROLOGY （ 影响因子：2.574； 五年影响因子：2.466 ）

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收录情况： SCI

摘要： The myristoylated capsid precursor mP1-2A of foot-and-mouth disease virus (FMDV), when expressed in mammalian cells and processed by the FMDV 3C protease, can self-assemble into virus-like particles (VLPs). In the present study, nine amino acids of the 3C protease were replaced by site-directed mutagenesis to create a mutant 3C protease, 9m3C. To coexpress mP1-2A and 9m3C and test the resulting proteolytic processing and VLP assembly, two recombinant goatpox viruses (rGTPVs) were constructed by the insertion of two coding regions, one for mP1-2A and the other for either 9m3C (rGTPV-mP1-2A-9m3C) or Theileria protective antigen (TPA) as a control (rGTPV-mP1-2A-TPA). The two exogenous genes were inserted into an intergenic region between loci gp_24 and gp_24.5 of the rGTPV genome. Western blotting of cells infected with rGTPV-mP1-2A-9m3C showed that proteins VP0, VP1, and VP3 from the mP1-2A processed by the 9m3C protease could be detected by polyclonal FMDV sera. As observed by electron microscopy, the infected cells produced VLPs with a diameter of about 25 ± 2 nm. Titers of neutralizing antibody against FMDV were significantly higher in mice inoculated with rGTPV-mP1-2A-9m3C, which expresses the 9m3C protease together with mP1-2A, than mice inoculated with the control rGTPV-mP1-2A-TPA, which does not express the protease. An ovine immunization test determined that sheep inoculated intramuscularly with rGTPV-mP1-2A-9m3C produced FMDV-specific neutralizing antibody, but its titers did not meet the requirement of the World Organization for Animal Health. The result indicates that further modifications of rGTPV-mP1-2A-9m3C are necessary to produce an effective vaccine.

分类号： R37

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