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Department of Biology, Biology Research Center, Imam Hosein University, Tehran , honari.hosein@gmail.com
Abstract:   (45 Views)
Background: One of the ways to strengthen the effect of vaccines is using the adjuvant. STxB has a carrier and adjuvant role that we can fuse it with vaccine candidate antigens and produce efficient vaccines. Saponaria Officinalis is a plant that has shown N-glycosidase activity. SO6 isoform of this plant, depurinate the adenine 4324 in the conserved sequence GAGA in 28SrRNA and disrupt protein synthesis. The aim of this study, was expression of SO6-STxB Gene in E. coli and investigation antibody titer in mice.
methods: In this study SO6 gene with BamHI and salI restriction enzyme sites were isolated from pUC57 plasmid and subcloned into expresson vector pET28a(+)-STxB and transformed into E. coli BL21 DE3. Expression of SO6-STxB gene Cassette was induced by IPTG and purified by nickel affinity chromatography. The recombinant protein was confirmed by western blotting. Mice were immunized intraperitoneally with purified protein and Serum IgG titers were measured by ELISA.
Results: subcloning of SO6–STxB gene in pET28a (+) expression vector confirmed by PCR and enzyme digestion reaction. A 37/5 kDa recombinant protein was confirmed by SDS-PAGE and western blotting. The antibodi generated from mouse serum isolated and was confirmed by ELISA.
Conclusion: Purified recombinant antigen STxB-SO6 can be used for research and anti-cancer vaccine candidate
Type of Study: Research | Subject: Biology

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