Volume 24, Issue 11 (Feb 2017)
JSSU 2017, 24(11): 876-886 |
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Masoudi Kerahroudi M, Honari H, abdollahi M. Expression of Blf1-Stx B Gene Cassette in E. coli and Investigation Antibody Titer in Mice. JSSU 2017; 24 (11) :876-886
URL: http://jssu.ssu.ac.ir/article-1-3921-en.html
URL: http://jssu.ssu.ac.ir/article-1-3921-en.html
Abstract: (6577 Views)
Background and Objectives: 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.
Methods: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.
Results: 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.
Conclusion: 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.
Type of Study: Original article |
Subject:
Genetics
Received: 2016/10/9 | Accepted: 2017/01/22 | Published: 2017/04/5
Received: 2016/10/9 | Accepted: 2017/01/22 | Published: 2017/04/5
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