Schistosoma japonicum gene recombination living vaccine production technique taking porcine pseudorabies virus as vector and product
A porcine pseudorabies and recombinant vaccine technology, applied in the field of production technology and products of Schistosoma japonicum gene recombinant live vaccine, can solve the problems of large schistosomiasis, unsatisfactory immune protection effect, complex antigen, etc., and achieve long shelf life and good immunity Protective effect, non-toxic side effects
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Embodiment 1
[0031] Preparation of Live Vaccine with Recombinant Schistosoma japonicum Sj26GST Gene Pseudorabies Virus Vector
[0032] According to attached figure 1 The process route shown is to prepare the recombinant Schistosoma japonicum Sj26GST gene pseudorabies virus vector live vaccine. Specific steps are as follows:
[0033] 1. Cloning of Schistosoma japonicum protective antigen Sj26GST gene and construction of eukaryotic expression vector
[0034] Rabbits were infected with 1500 cercariae of Schistosoma japonicum and dissected after 45 days. Perfuse the portal vein with sterile sodium citrate saline, collect adult worms, and wash with sterile saline to remove blood components attached to the surface of the worms. Take 0.5 g of fresh adults, homogenate on ice, and use the Trizol method to prepare adult mRNA. The protective antigen Sj26GST was obtained by RT-PCR. After the amplified product was confirmed by agarose gel electrophoresis, the obtained gene was cloned into the pMD-...
Embodiment 2
[0054] Preparation of Live Vaccine with Recombinant Schistosoma japonicum Sj23 Gene Pseudorabies Virus Vector
[0055] According to attached figure 1 The process route shown is to prepare the recombinant Schistosoma japonicum Sj23 gene pseudorabies virus vector live vaccine. Specific steps are as follows:
[0056] 1. Cloning of Schistosoma japonicum protective antigen Sj23 gene and construction of eukaryotic expression vector
[0057] Rabbits were infected with 1500 cercariae of Schistosoma japonicum and dissected after 45 days. Perfuse the portal vein with sterile sodium citrate saline, collect adult worms, and wash with sterile saline to remove blood components attached to the surface of the worms. Take 0.5 g of fresh adults, homogenate on ice, and use the Trizol method to prepare adult mRNA. The protective antigen Sj23 was obtained by RT-PCR. After the amplified product was confirmed by agarose gel electrophoresis, the obtained gene was cloned into the pMD-18T vector t...
Embodiment 3
[0077] Preparation of Live Vaccine with Recombinant Schistosoma japonicum FABP Gene and Pseudorabies Virus Vector
[0078] According to attached figure 1 The shown process route prepares the recombinant schistosoma japonicum FABP gene pseudorabies virus vector live vaccine. Specific steps are as follows:
[0079] 1. Cloning of Schistosoma japonicum protective antigen FABP gene and construction of eukaryotic expression vector
[0080] Rabbits were infected with 1500 cercariae of Schistosoma japonicum and dissected after 45 days. Perfuse the portal vein with sterile sodium citrate saline, collect adult worms, and wash with sterile saline to remove blood components attached to the surface of the worm body. Take 0.5 g of fresh adults, homogenate on ice, and use the Trizol method to prepare adult mRNA. The protective antigen FABP was obtained by RT-PCR. After the amplified product was confirmed by agarose gel electrophoresis, the obtained gene was cloned into the pMD-18T vector ...
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