Expression of Blf1-Stx B Gene Cassette in E. coli and Investigation Antibody Titer in Mice
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 pseudomall...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | fas |
| Published: |
Shahid Sadoughi University of Medical Sciences
2017-02-01
|
| Series: | Majallah-i Dānishgāh-i ’Ulūm-i Pizishkī-i Shahīd Ṣadūqī Yazd |
| Subjects: | |
| Online Access: | http://jssu.ssu.ac.ir/browse.php?a_code=A-10-2958-1&slc_lang=en&sid=1 |
| Summary: | 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. |
|---|---|
| ISSN: | 2228-5741 2228-5733 |