Establishment of <i>Echinococcus granulosus EgM123</i> Recombinant Gene Rabies Virus SRV<sub>9</sub> and Identification of Its Biological Characteristics
Canids act as a crucial intermediary in the transmission of rabies and <i>Echinococcus granulosus</i>, serving as co-infection hosts and pathogen carriers for both rabies and hydatid disease (HD) transmitted from animals to humans. Therefore, an effective and efficient bivalent oral vacc...
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| author | Yueqi Yang Mengdan Hou Guicheng Su Xiaoyan Ma Xiaohui Su Kunlei Li Songhan Liu Luheng Xiao Jingjing Yao Jiahao Zhai Xiaoying Wei Yang Zhou Qianqian Lai Yuwei Dong Jieyu Liu Shaohua Zhai |
| author_facet | Yueqi Yang Mengdan Hou Guicheng Su Xiaoyan Ma Xiaohui Su Kunlei Li Songhan Liu Luheng Xiao Jingjing Yao Jiahao Zhai Xiaoying Wei Yang Zhou Qianqian Lai Yuwei Dong Jieyu Liu Shaohua Zhai |
| author_sort | Yueqi Yang |
| collection | DOAJ |
| description | Canids act as a crucial intermediary in the transmission of rabies and <i>Echinococcus granulosus</i>, serving as co-infection hosts and pathogen carriers for both rabies and hydatid disease (HD) transmitted from animals to humans. Therefore, an effective and efficient bivalent oral vaccine for preventing HD and rabies is urgently required to reduce economic losses in husbandry resulting from rabies and HD. In this study, a full-length plasmid (pcDNA4-NPM+G<sub>ΔCD</sub>+EgM123+eGFP+L) carrying the <i>Echinococcus granulosus EgM123</i> gene and fluorescence reporter genes of eGFP and four auxiliary transfection plasmids of rabies virus SRV<sub>9</sub> (pcDNA4-N, pcDNA4-P, pcDNA4-G, pcDNA-L) were established by reverse genetics approaches and co-transfected to BSR cells by electrotransfection. The co-transfected BSR cells showed green fluorescence 48 h after electrotransfection. The recombinant virus was exposed to the sixth-generation blind passage, with the <i>N</i>, <i>P</i>, <i>G</i>, and <i>EgM123</i> genes amplified via RT-PCR, yielding targeted strips. The rescued virus-infected BSR cells were characterized by TEM, and the results indicated that bullet-like viral particles with an average size of 148.47 nm and a cyst structure were present in the cytoplasm of BSR cells; the expression levels of continuously cultivated 9th-, 10th-, 11th-, 12th-, and 13th-generation viruses were quantified by qRT-PCR, and the results showed that mRNA expression of the virus was upregulated. The LD<sub>50</sub> titer of suckling rats was measured to be 10<sup>−1.4</sup>. The synthesized <i>EgM123</i> recombinant gene rabies virus SRV<sub>9</sub> can function as a vaccine strain for the development of the “Rabies-HD bivalent recombinant gene oral vaccine”, therefore aiding in the prevention and management of rabies and HD in animals. |
| format | Article |
| id | doaj-art-74979cd713014f07bc0ae0ae2dbd85ae |
| institution | Kabale University |
| issn | 1999-4915 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
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| series | Viruses |
| spelling | doaj-art-74979cd713014f07bc0ae0ae2dbd85ae2025-01-24T13:52:19ZengMDPI AGViruses1999-49152024-12-011713010.3390/v17010030Establishment of <i>Echinococcus granulosus EgM123</i> Recombinant Gene Rabies Virus SRV<sub>9</sub> and Identification of Its Biological CharacteristicsYueqi Yang0Mengdan Hou1Guicheng Su2Xiaoyan Ma3Xiaohui Su4Kunlei Li5Songhan Liu6Luheng Xiao7Jingjing Yao8Jiahao Zhai9Xiaoying Wei10Yang Zhou11Qianqian Lai12Yuwei Dong13Jieyu Liu14Shaohua Zhai15School of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaLivestock and Veterinary Work Station of Xinjiang Production and Construction Corps, Urumqi 830063, ChinaLivestock and Veterinary Work Station of Xinjiang Production and Construction Corps, Urumqi 830063, ChinaLivestock and Veterinary Work Station of Xinjiang Production and Construction Corps, Urumqi 830063, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaSchool of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, ChinaCanids act as a crucial intermediary in the transmission of rabies and <i>Echinococcus granulosus</i>, serving as co-infection hosts and pathogen carriers for both rabies and hydatid disease (HD) transmitted from animals to humans. Therefore, an effective and efficient bivalent oral vaccine for preventing HD and rabies is urgently required to reduce economic losses in husbandry resulting from rabies and HD. In this study, a full-length plasmid (pcDNA4-NPM+G<sub>ΔCD</sub>+EgM123+eGFP+L) carrying the <i>Echinococcus granulosus EgM123</i> gene and fluorescence reporter genes of eGFP and four auxiliary transfection plasmids of rabies virus SRV<sub>9</sub> (pcDNA4-N, pcDNA4-P, pcDNA4-G, pcDNA-L) were established by reverse genetics approaches and co-transfected to BSR cells by electrotransfection. The co-transfected BSR cells showed green fluorescence 48 h after electrotransfection. The recombinant virus was exposed to the sixth-generation blind passage, with the <i>N</i>, <i>P</i>, <i>G</i>, and <i>EgM123</i> genes amplified via RT-PCR, yielding targeted strips. The rescued virus-infected BSR cells were characterized by TEM, and the results indicated that bullet-like viral particles with an average size of 148.47 nm and a cyst structure were present in the cytoplasm of BSR cells; the expression levels of continuously cultivated 9th-, 10th-, 11th-, 12th-, and 13th-generation viruses were quantified by qRT-PCR, and the results showed that mRNA expression of the virus was upregulated. The LD<sub>50</sub> titer of suckling rats was measured to be 10<sup>−1.4</sup>. The synthesized <i>EgM123</i> recombinant gene rabies virus SRV<sub>9</sub> can function as a vaccine strain for the development of the “Rabies-HD bivalent recombinant gene oral vaccine”, therefore aiding in the prevention and management of rabies and HD in animals.https://www.mdpi.com/1999-4915/17/1/30rabies virus SRV<sub>9</sub><i>Echinococcus granulosus EgM123</i> genereverse geneticsvirus rescue |
| spellingShingle | Yueqi Yang Mengdan Hou Guicheng Su Xiaoyan Ma Xiaohui Su Kunlei Li Songhan Liu Luheng Xiao Jingjing Yao Jiahao Zhai Xiaoying Wei Yang Zhou Qianqian Lai Yuwei Dong Jieyu Liu Shaohua Zhai Establishment of <i>Echinococcus granulosus EgM123</i> Recombinant Gene Rabies Virus SRV<sub>9</sub> and Identification of Its Biological Characteristics Viruses rabies virus SRV<sub>9</sub> <i>Echinococcus granulosus EgM123</i> gene reverse genetics virus rescue |
| title | Establishment of <i>Echinococcus granulosus EgM123</i> Recombinant Gene Rabies Virus SRV<sub>9</sub> and Identification of Its Biological Characteristics |
| title_full | Establishment of <i>Echinococcus granulosus EgM123</i> Recombinant Gene Rabies Virus SRV<sub>9</sub> and Identification of Its Biological Characteristics |
| title_fullStr | Establishment of <i>Echinococcus granulosus EgM123</i> Recombinant Gene Rabies Virus SRV<sub>9</sub> and Identification of Its Biological Characteristics |
| title_full_unstemmed | Establishment of <i>Echinococcus granulosus EgM123</i> Recombinant Gene Rabies Virus SRV<sub>9</sub> and Identification of Its Biological Characteristics |
| title_short | Establishment of <i>Echinococcus granulosus EgM123</i> Recombinant Gene Rabies Virus SRV<sub>9</sub> and Identification of Its Biological Characteristics |
| title_sort | establishment of i echinococcus granulosus egm123 i recombinant gene rabies virus srv sub 9 sub and identification of its biological characteristics |
| topic | rabies virus SRV<sub>9</sub> <i>Echinococcus granulosus EgM123</i> gene reverse genetics virus rescue |
| url | https://www.mdpi.com/1999-4915/17/1/30 |
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