Construction and Immogenecity of Pseudorabies Virus CY-6 strain with gE/gI Deletion
BHK-21 cells were co-transfected with porcine pseudorabies virus PRV-CY-ΔgI/gE-GFP,pBL-gI-gE vector and Lipofectamin2000 transfection agent to construct a recombinant virus strain PRV-CY-ΔgI/gE.The CY-6 strain was subject to stain screening and plague purification for further study.Subsequently,BHK-21 cells were inoculated with the CY-6 strain from C1 to C10 passages and the virus harvests were tested for their growth curves,virus contents,gene identification,virulence,purity,specificity and immunogenicity.Results showed that the growth curve of CY-6 Strain had no significant change from its parent CY.The virus contents of C1-C10 passages ranged from 108.17 to 108.68 TCID50/mL.Gene identification showed that the CY-6 had gE and g/I genes but possessed gD gene of a 217 bp target band.Purity test demonstrated its sterility for mycoplasma and foreign virus contamination.In addition,the CY-6 was neutralized by specific positive PRV antiserum.In the immunogenicity study,5 pigs of the CY-6 immunized group was all protected and 5 pigs of the challenge control group was infected and 3 pigs died.These results indicated that the CY-6 strain fulfilled the preliminary requirements for the production of virus strains,which laid the foundation for the development of a novel inactivated,gene-deleted PRV vaccine.
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