Volume 25, Issue 11 (2-2019)                   RJMS 2019, 25(11): 52-60 | Back to browse issues page

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abdollahi M, Honari H, e'temad aubi S M. Expression of SO6-STxB gene cassette in Escherichia coli and investigation of antibody titer. RJMS. 2019; 25 (11) :52-60
URL: http://rjms.iums.ac.ir/article-1-5164-en.html
Imam Hosein University, Tehran Iran , honari.hosein@gmail.com
Abstract:   (197 Views)
Background: One of the ways to strengthen the effect of vaccines is the use of adjuvant. STxB has a carrier role and can act as an adjuvant; thus it can be fused with vaccine candidate antigens in order to produce efficient vaccines. Saponaria officinalis is a plant that has shown N-glycosidase activity. SO6 isoform of this plant, depurinates the adenine 4324 in the conserved sequence GAGA in 28SrRNA and disrupts protein synthesis. The aim of this study was expression of SO6-STxB gene in Escherichia coli (E. coli) and investigation of 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 pET28a (+) -STxB expression vector and transferred to 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 was confirmed by PCR and enzyme digestion reaction. A 37/5 kDa recombinant protein was confirmed by SDS-PAGE and western blotting. The antibody generated from mouse serum was isolated and confirmed by ELISA.
Conclusion: Purified recombinant antigen STxB-SO6 can be used for research and be a suitable anti-cancer vaccine candidates.
 
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Type of Study: Research | Subject: Biology

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