Schistosoma japonicum thioredoxin glutathione reductase gene and DNA (deoxyribonucleic acid) vaccine thereof
A technology of thioredoxin and glutathione, applied in the field of bioengineering, can solve problems such as reinfection that cannot be solved
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Embodiment 1
[0021] Example 1 Cloning of Schistosoma japonicum Thioredoxin Glutathione Reductase SjTGR Gene and Construction of SjTGR-pVAX1 Recombinant Plasmid
[0022] 1.1 Experimental animals Male New Zealand white rabbits, weighing 2.5kg to 3kg, were purchased from Shanghai Luojing Feida Experimental Animal Farm.
[0023] 1.2 Adult Schistosoma japonicum (strain from mainland China) The cercariae of Schistosoma japonicum were provided by the snail room of Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences. New Zealand white rabbits were infected with 2000 cercariae of Schistosoma japonicum by abdominal patch method, and were dissected 42 days after infection. Adult worms of Schistosoma japonicum were collected from the hepatic portal vein by thoracic aorta perfusion method, and the worms were fully washed with sterile PBS to remove the adhesion from the host. substance. The washed adult schistosomes were frozen in liquid nitrogen for later use.
[0024] 1....
Embodiment 2
[0056] Example 2 Evaluation of the immune protection effect of the recombinant plasmid SjTGR-pVAX1 in the mouse schistosomiasis model
[0057] 1. Materials
[0058] Male BALB / c mice aged 5-6 weeks were purchased from the Department of Experimental Animal Science, Fudan University. The cercariae of Schistosoma japonicum strains from mainland China were provided by the Snail Laboratory of Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences.
[0059] Reagents and equipment related to the gene gun were purchased from bio-rad company, and the operation was also carried out according to its instructions. Other reagents are the same as above.
[0060] 2. DNA vaccine preparation and immunization methods
[0061] (1) First determine that the amount of metal particle carrier (MLQ) is 0.5 mg / bullet, and the loading ratio (DLR) of DNA is 5 μg / mg gold powder.
[0062] (2) Coating DNA onto the metal particle carrier.
[0063] 1) Take the PVP (Polyvinylpyrro...
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