Pseudorabies virus gene engineering gB recombinant attenuated vaccine strain and application thereof
A technology of pseudorabies virus and attenuated vaccine, which is applied in the genetically engineered gB recombinant attenuated vaccine strain of pseudorabies virus and its application field, can solve the problems of inability to resist the infection of porcine pseudorabies mutant strains, and achieve prevention and control, good immunogen sexual effect
Active Publication Date: 2017-07-07
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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[0004] In order to solve the problems such as the porcine pseudorabies vaccine can not resist the porcine pseudorabies mutant strain infection in the above prior art, the invention provides a kind of pseudorabies virus genetic engineering gB recombinant virus Bar-JSgB (BJB) that can be used to prepare the vaccine with high immune efficiency ), the vaccine strain is based on the classic pseudorabies virus attenuated vaccine strain Bartha-K61, and its main protective antigen gB gene coding region is replaced by the gB gene coding region of the pseudorabies mutant strain JS-2012, and the Bartha-K61 The attenuated characteristics of JS-2012 gB protein combined with the immunogenicity of the mutant strain JS-2012 gB protein, the obtained recombinant strain
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Embodiment 2
[0069] Example 2 Recombinant PRV Attenuated Vaccine Strain Inactivated Vaccine Immune Protection Test
[0070] In this example, the recombinant PRV virus Bar-JSgB and the vaccine strain Bartha-K61 were respectively used as seed strains to prepare inactivated vaccines, and after the mice were immunized, the virulent strain JS-2012 was used for challenge. ) inactivated vaccine immunization group had a better protective effect than Bartha-K61 inactivated vaccine immunization group. The specific process is as follows:
[0071] 2.1 Preparation of recombinant PRV attenuated vaccine strain inactivated vaccine
[0072] The recombinant PRV attenuated vaccine strain Bar-JSgB (BJB) obtained in Example 1 and the parental strain Bartha-K61 were respectively inoculated into PK-15 cells for amplification. After the cells were completely damaged, the supernatant of the cells was collected and centrifuged at a high speed, and retained. Supernatant, remove cell debris. Formaldehyde was mixed...
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The invention discloses a pseudorabies virus gene engineering gB recombinant attenuated vaccine strain and application thereof. CRISPR/Cas9 is combined with techniques such as homologous recombination, and a genome of a pseudorabies virus vaccine strain Bartha-K61 is taken as a framework, and a recombinant pseudorabies virus strain Bar-JS-gB (BJB) is successfully acquired by replacing a gB gene of the Bartha-K61 with a gB gene of an epidemic pseudorabies virus variant JS-2012 in China. The strain BJB has stable heredity, and an immune mouse processed by an inactivated vaccine prepared by taking the strain BJB as a vaccine strain has relatively good capacity of effectively resisting attacking of a virulent strain JS-2012 than an immune mouse processed by an inactivated vaccine prepared from the Bartha-K61. Therefore, the strain BJB has relatively good immunogenicity, can be used for effectively controlling the prevalence of a PRV variant in China and can play an important role on control of pseudorabies.
Description
technical field [0001] The invention relates to the field of microbes, in particular to the technical field of pseudorabies virus, in particular to a genetically engineered gB recombinant attenuated vaccine strain of pseudorabies virus and its application. Background technique [0002] Porcine pseudorabies (Pseudorabies, PR) is one of the main diseases that seriously endanger the pig industry in my country. The disease is caused by pseudorabies virus (Pseudorabies Virus, PRV) infection. Its main clinical symptoms are typical nervous system diseases, It is characterized by high fever, severe itching and dyspnea, reproductive disorders such as stillbirth and mummified fetuses in sows, and high lethality in piglets. PRV belongs to the herpesviridae herpesvirus subfamily A, its genome is a linear double-stranded DNA, about 150kb in length, composed of a long unique region (UL), a short unique region (US) and repeat sequences on both sides of the US, encoding There are more than ...
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IPC IPC(8): C12N7/01C12N15/38A61K39/245A61P31/22C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/552C07K14/005C12N7/00C12N2710/16721C12N2710/16722C12N2710/16734
Inventor 童光志于之清童武郑浩李国新
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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