Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus
Peste des petits ruminants (PPR) is one of the most contagious and fatal diseases of small ruminants in the world and is classified as a category A epidemic disease. It is the target of a global eradication campaign led by the Office International des Epizooties (OIE) and Food and Agriculture Organi...
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| Format: | Article |
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MDPI AG
2022-10-01
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| Series: | Veterinary Sciences |
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| Online Access: | https://www.mdpi.com/2306-7381/9/10/555 |
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| author | Haojie Wang Jinhao Bi Na Feng Yongkun Zhao Tiecheng Wang Yuetao Li Feihu Yan Songtao Yang Xianzhu Xia |
| author_facet | Haojie Wang Jinhao Bi Na Feng Yongkun Zhao Tiecheng Wang Yuetao Li Feihu Yan Songtao Yang Xianzhu Xia |
| author_sort | Haojie Wang |
| collection | DOAJ |
| description | Peste des petits ruminants (PPR) is one of the most contagious and fatal diseases of small ruminants in the world and is classified as a category A epidemic disease. It is the target of a global eradication campaign led by the Office International des Epizooties (OIE) and Food and Agriculture Organization of the United Nations (FAO). The PPR live attenuated vaccine is currently the most widely used and approved vaccine, but the use of this vaccine interferes with the serological testing of the PPR elimination program, and there is a potential safety risk. Viral vector vaccines are one of the most promising methods to solve this dilemma. In this study, the full-length infectious clone plasmid of rabies virus (RABV), pD-SRV9-PM-LASV, was used as the backbone, and the envelope glycoprotein H (hemagglutinin protein) or F (fusion protein) gene of PPRV was inserted into the backbone plasmid to construct the infectious clones pD-SRV9-PM-PPRV-H and pD-SRV9-PM-PPRV-F, which express the PPRV H and PPRV F genes, respectively. The correct construction of these infectious clones was verified after sequencing and double digestion. The infectious clones were transfected with a helper plasmid into BSR/T7 cells, and recombinant viruses were successfully rescued by direct immunofluorescence, indirect immunofluorescence, Western blotting, and transmission electron microscopy and named rSRV9-H and rSRV9-F. The results of growth kinetics studies indicated that the inserted gene did not affect virus proliferation. Stability studies revealed that the inserted target gene was stably expressed in recombinant RABV for at least 15 generations. In this study, the recombinant viruses rSRV9-H and rSRV9-F were successfully rescued. The constructed viruses had good proliferative activity and stability and provided potential bivalent inactivated vaccine candidate strains for the prevention of PPR and livestock rabies. |
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| institution | Directory Open Access Journal |
| issn | 2306-7381 |
| language | English |
| last_indexed | 2024-03-09T19:24:18Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Veterinary Sciences |
| spelling | doaj.art-cadfd27595884bf38bdf04e5c02795922023-11-24T03:06:36ZengMDPI AGVeterinary Sciences2306-73812022-10-0191055510.3390/vetsci9100555Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants VirusHaojie Wang0Jinhao Bi1Na Feng2Yongkun Zhao3Tiecheng Wang4Yuetao Li5Feihu Yan6Songtao Yang7Xianzhu Xia8Changchun Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Changchun 130000, ChinaCollege of Veterinary Medicine, Jilin Agriculture University, Changchun 453003, ChinaChangchun Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Changchun 130000, ChinaChangchun Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Changchun 130000, ChinaChangchun Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Changchun 130000, ChinaCollege of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang 130118, ChinaChangchun Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Changchun 130000, ChinaChangchun Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Changchun 130000, ChinaChangchun Veterinary Research Institute, Chinese Academy of Agriculture Sciences, Changchun 130000, ChinaPeste des petits ruminants (PPR) is one of the most contagious and fatal diseases of small ruminants in the world and is classified as a category A epidemic disease. It is the target of a global eradication campaign led by the Office International des Epizooties (OIE) and Food and Agriculture Organization of the United Nations (FAO). The PPR live attenuated vaccine is currently the most widely used and approved vaccine, but the use of this vaccine interferes with the serological testing of the PPR elimination program, and there is a potential safety risk. Viral vector vaccines are one of the most promising methods to solve this dilemma. In this study, the full-length infectious clone plasmid of rabies virus (RABV), pD-SRV9-PM-LASV, was used as the backbone, and the envelope glycoprotein H (hemagglutinin protein) or F (fusion protein) gene of PPRV was inserted into the backbone plasmid to construct the infectious clones pD-SRV9-PM-PPRV-H and pD-SRV9-PM-PPRV-F, which express the PPRV H and PPRV F genes, respectively. The correct construction of these infectious clones was verified after sequencing and double digestion. The infectious clones were transfected with a helper plasmid into BSR/T7 cells, and recombinant viruses were successfully rescued by direct immunofluorescence, indirect immunofluorescence, Western blotting, and transmission electron microscopy and named rSRV9-H and rSRV9-F. The results of growth kinetics studies indicated that the inserted gene did not affect virus proliferation. Stability studies revealed that the inserted target gene was stably expressed in recombinant RABV for at least 15 generations. In this study, the recombinant viruses rSRV9-H and rSRV9-F were successfully rescued. The constructed viruses had good proliferative activity and stability and provided potential bivalent inactivated vaccine candidate strains for the prevention of PPR and livestock rabies.https://www.mdpi.com/2306-7381/9/10/555peste des petits ruminantsrabies virus vectorreverse geneticsvaccine candidate strainbivalent vaccine |
| spellingShingle | Haojie Wang Jinhao Bi Na Feng Yongkun Zhao Tiecheng Wang Yuetao Li Feihu Yan Songtao Yang Xianzhu Xia Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus Veterinary Sciences peste des petits ruminants rabies virus vector reverse genetics vaccine candidate strain bivalent vaccine |
| title | Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus |
| title_full | Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus |
| title_fullStr | Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus |
| title_full_unstemmed | Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus |
| title_short | Construction of Recombinant Rabies Virus Vectors Expressing H or F Protein of Peste des Petits Ruminants Virus |
| title_sort | construction of recombinant rabies virus vectors expressing h or f protein of peste des petits ruminants virus |
| topic | peste des petits ruminants rabies virus vector reverse genetics vaccine candidate strain bivalent vaccine |
| url | https://www.mdpi.com/2306-7381/9/10/555 |
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