Study On Immunoprotection Of Neutral Protease Against Anthrax

People and livestock infected by Bacillus anthraris which causes acute and fierce epidemic infectious disease, because one of this kind of disease symptom types is causes organizations and skin have the black coal shape necrosis,Therefore it is called"anthrax". This disease spread all over the world nearly, farmer, herdsman and animal slaughter as well as raising livestock veterinarian are all easiest to infect. Due to the recombinant PA vaccine is lower than other vaccines with the protection ratio, Therefore,Only depended on the PA antigen become unreasonable. Scientists think that had other protective antigen besides PA.First, through had the protective function antigen ingredient to the Chinese anthrax PA vaccine to carry on the qualitative analysis, the protective antigen ingredient through the high-pressured liquid phase purification confirmation molecular weight was 32 ku proteins,Since this value is lower than the PA molecular weight which has recognized continuously (83 ku).We carry on SDS-PAGE for the Chinese anthrax PA vaccine,Cutting the protective molecular band to carry on the protein fingerprint atlas to analyze (MALDI-TOF-MS). According to MALDI-TOF result,The protein determination for anthrax neutral protease (neutral protease,NPR), but is not PA.Second, the object of study selected Bacillus anthraris PA,NPR and the neutral protease conservative position code gene.PCR clone Bacillus anthraris PA, N and NPR code gene, after cloning connect with pGEM-T, after that we sequencing the PCR proudct and connectting with the prokaryotic expressing vector pET28a(+) and pQE80-L,constructs the recombinant prokaryotic expressing plasmids pET28a-PA,pET28a-N and pQE80-NPR. Through IPTG induction expression,The SDS-PAGE detect the express product, Western Blotting method appraisal result demonstrated the recombinant plasmids can express PA, NPR and the N protein.Third,the recombinant protein after purification add-on Al(OH)3 adjuvant immunity Balb/c mouse,the examination cellular immunity and the humoral immunity target, in the ELISA method examination mouse blood serum the total IgG horizontal...