Recombinant porcine pseudorabies virus strain used for expression of porcine circovirus type II (PCV2) ORF2 gene, and preparation method thereof

Abstract

The invention discloses a recombinant porcine pseudorabies virus (Herpesviridae) strain used for expression of porcine circovirus type II ORF2 gene. Preservation number of the recombinant porcine pseudorabies virus strain is CCTCC No.V201315. According to the preparation method, PCV2ORF2 gene is inserted into common carrier PG so as to construct transferring plasmid PGO; monolayer ST cells are inoculated with PRVTK<-> / gG<-> / gE<-> virus by 2h of adsorption; ST cells are transfected with plasmid PGO, wherein fusion degree of the monolayer ST cells is 80 to 90%; the recombinant porcine pseudorabies virus PGO strain is obtained by plaque purification, and is used for immunization of mouse. It is shown by results that commercial PCV2 inactivated vaccine and a group immunized by the recombinant porcine pseudorabies virus PGO strain are both capable of inducing specific humoral immune response of mouse, antibody titers of both the commercial PCV2 inactivated vaccine and the group are obviously higher than that of a group immunized by DMEM medium, and difference is significant (p<0.05). It is shown by the results of mouse lymphocyte proliferation test that specific ceullar immune response caused by the group immunized by the recombinant porcine pseudorabies virus PGO strain is more obvious than that caused by the commercial PCV2 inactivated vaccine and the group immunized by DMEM medium, and difference is obvious (p<0.05). In addition, the group immunized by the recombinant porcine pseudorabies virus PGO strain is capable of resisting severe attack by PCV2. Therefore application of the recombinant porcine pseudorabies virus strain in development of a novel PCV2 vaccine is possible.

Description

technical field [0001] The invention relates to a method for preparing a recombinant porcine pseudorabies virus PGO strain expressing porcine circovirus type II ORF2 gene. Background technique [0002] Porcine circovirus (Porcine circovirus, PCV) belongs to the family Circoviridae and is the smallest vertebrate virus discovered so far. PCV is divided into two genotypes, PCV1 and PCV2, according to the pathogenicity, antigenicity and nucleotide sequence. Among them, PCV1 widely exists in pigs but has no pathogenicity to it, and its genome length is 1759bp. PCV2 is pathogenic, with a gene length of 1767bp or 1768bp. PCV2 is not only the pathogenic agent of post-weaning multisystemic failure syndrome (PMWS) in piglets, but also associated with porcine dermatitis and nephrotic syndrome (PDNS), sow Abortion, swine reproductive failure, pig inflammatory respiratory disease syndrome (PRDC) and mortality syndrome (SAMA) have an important relationship. In addition, PCV2 can infrin...

AI Technical Summary

Problems solved by technology

In addition, PCV2 can infringe on the lymphatic system to cause immunosuppression in pigs, and secondary infection of swine flu, porcine pseudorabies, porcine reproductive and respiratory syndrome, swine fever and other viral infectious diseases and various bacterial infectious diseases, thus making the epidemic situation become more severe. It is more serious. The widespread existence of the virus at this stage has caused serious economic losses to the pig industry in my country and other countries around the world.

At present, there are few vaccines that can effectively prevent and control this disease at home and abroad. Therefore, research on safe and effective vaccines for PCV2 is of great significance for the control and eradication of PCV2-related diseases.

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