Recombinant new castle disease LaSota attenuated vaccine strain expressing rabies virus glucoprotein (GP protein)
A technology of rabies virus and attenuated vaccines, which is applied in the field of recombinant virus vaccines and can solve problems such as limitations in the scope of use
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Embodiment 1
[0036] Example 1 Construction and biological activity of a recombinant Newcastle disease LaSota attenuated vaccine expressing a gene encoding rabies virus glycoprotein (GP protein)
[0037] 1 Materials and methods
[0038] 1.1 Cells and viruses
[0039] BHK-21 cells (baby hamster kidney cells ATCC CCL-10), Vero-E6 cells (African green monkey kidney cells ATCC No.CRL-1586), the medium is DMEM with 5% fetal bovine serum; rabies virus (RV) ERA strain (AV61) was purchased from the China Institute of Veterinary Drug Control, and the virus titer was titrated on Vero-E6 cells; the recombinant poxvirus vTF7-3 (ATCC, VR-2153) stably expressing T7 polymerase was purchased from ATCC; ERA immunization Mouse serum and chicken anti-NDV hyperimmune serum were purchased from China Veterinary Drug Supervision Institute.
[0040] 1.2 Main reagents and instruments
[0041] FITC-labeled rabbit anti-chicken fluorescent secondary antibody and anti-mouse fluorescent secondary antibody were purchased fro...
Embodiment 2
[0064] Example 2 Immune test of recombinant virus to BALB / c mice
[0065] 1. Materials and methods
[0066] 1.1 Material
[0067] Same as Example 1 above.
[0068] 1.2 Method
[0069] Immune test of recombinant virus to BALB / c mice
[0070] In order to evaluate the immune effect of the recombinant virus on BALB / c mice, the chicken embryos of Example 1 were amplified with allantoxin to 2x10 8 EID 50 Dose Artificial immunization of 6-week-old BALB / c mice (purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.) via intraperitoneal injection, and 8 non-immune control groups; the immunized group and the non-immune control group were raised separately In the air negative pressure filter isolator. 21 days after immunization, blood was collected from the heart and the serum was separated according to routine tests for NDV-specific hemagglutination inhibition (HI) antibodies. The virus neutralization test was performed with ERA as the virus antigen to detect anti-ERA a...
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