Expression of Blf1-Stx B Gene Cassette in E. coli and Investigation Antibody Titer in Mice

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. BLF1 has a role in the pathogenesis and infection by Burkholderia pseudomallei, that can be studied when fused with stxB.The aim of this study expression blf1-stxB gene Cassette in E. coli and antibody production in mice.
In this study the synthetic gene in pUC57 plasmid was purchased from Nedaye Fan COR. pUC57 plasmid containing blf1 with BamHI , SalI restriction enzyme sites was subcloned in pET28a()-stxB expression vector and transformed into E. coli BL21 DE3. expression blf1-stxB gene Cassette was induced by IPTG. After extraction by affinity chromatography, the recombinant protein was injected four times to mice.
In this study, blf1-StxB cloned gene in pET28a() expression vector was approved by PCR and enzymatic analysis. Also recombinant protein confirmed by SDS-PAGE and Western blotting. Then antibody produced from the mice serum, were isolated and confirmed by ELISA.
Given that BLF1 protein has the ability to stop protein synthesis and STxB has a carrier and adjuvant role, also with respect to antibody titers produced, it’s as a vaccine candidate against B. pseudomallei and Shigella dysenteriae.