Volume 24, Number 11 (Feb 2017)                   JSSU 2017, 24(11): 876-886 | Back to browse issues page

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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

Abstract:   (1065 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.

Full-Text [PDF 704 kb]   (306 Downloads)    
Type of Study: Original article | Subject: Genetics
Received: 2016/10/9 | Accepted: 2017/01/22 | Published: 2017/04/5

1. Kanaphun P, Thirawattanasuk N, Suputtamongkol Y, Naigowit P, Dance DA, Smith MD, et al. Serology and carriage of Pseudomonas pseudomallei: a prospective study in 1000 hospitalized children in northeast Thailand. Journal of Infectious Diseases 1993; 167(1): 13-230.
2. Meselson M, Guillemin J, Hugh-Jones M. Public health assessment of potential biological terrorism agents. Emerging infectious diseases. 2002;8(2):225.
3. Lee YH, Chen Y, Ouyang X, Gan Y-H. Identification of tomato plant as a novel host model for Burkholderia pseudomallei. BMC microbiology 2010; 10(1): 1.
4. Koponen MA, Zlock D, Palmer DL, Merlin TL. Melioidosis: forgotten, but not gone! Archives of internal medicine 1991; 151(3): 605-8.
5. Currie BJ, Fisher DA, Howard DM, Burrow JN, Selvanayagam S, Snelling PL, et al. The epidemiology of melioidosis in Australia and Papua New Guinea. Acta tropica 2000; 74(2): 121-7.
6. Lee S-H, Chong C-E, Lim B-S, Chai S-J, Sam K-K, Mohamed R, et al. Burkholderia pseudomallei animal and human isolates from Malaysia exhibit different phenotypic characteristics. Diagnostic Microbiology and Infectious Disease 2007; 58(3): 263-70.
7. Holden MT, Titball RW, Peacock SJ, Cerdeño-Tárraga AM, Atkins T, Crossman LC, et al. Genomic plasticity of the causative agent of melioidosis, Burkholderia pseudomallei. Proceedings of the National Academy of Sciences of the United States of America 2004; 101(39): 14240-5.
8. Pause A, Methot N, Svitkin Y, Merrick W, Sonenberg N. Dominant negative mutants of mammalian translation initiation factor eIF-4A define a critical role for eIF-4F in cap-dependent and cap-independent initiation of translation. The EMBO journal 1994; 13(5): 1205.
9. Bouter A, Delord B, Dransart E, Poirier C, Johannes L, Effenterre D. Intracellular trafficking of Shiga‐toxin‐B‐subunit‐functionalized spherulites. Biology of the Cell 2008; 100(12): 717-28.
10. Pina DG, Johannes L. Cholera and Shiga toxin B-subunits: thermodynamic and structural considerations for function and biomedical applications. Toxicon 2005; 45(4): 93-308.
11. Janssen K-P, Vignjevic D, Boisgard R, Falguières T, Bousquet G, Decaudin D, et al. In vivo tumor targeting using a novel intestinal pathogen-based delivery approach. Cancer Research 2006; 66(14): 7230-36.
12. Honari H, Amlashi I, Minaee ME, Safaee S. Immunogenicity in guinea pigs by IpaD-STxB recombinant protein. Arak Medical University Journal. 2013;16(4):83-93. (Persian)
13. Honari H, Minaei HH, Ebrahim M. Analyzing the Various Fusions for ctxB, ipaD and stxB Genes of Shigella Dysenteriae and Vibrio Cholera by Bioinformatics Tools. Genetics in the 3rd Millennium 2013; 11(2): 3070-77. [Persian]
14. Sambrook J, Russell DW. Molecular cloning: a laboratory manual 3rd edition. Coldspring-Harbour Laboratory Press, UK. 2001.
15. Tonello F, Pellizzari R, Pasqualato S, Grandi G, Peggion E, Montecucco C. Recombinant and truncated tetanus neurotoxin light chain: cloning, expression, purification, and proteolytic activity. Protein Expression and Purification 1999; 15(2): 221-27.
16. Clemens J, Kotloff K, Kay BA. Generic protocol to estimate the burden of Shigella diarrhoea and dysenteric mortality: Citeseer; 1999.
17. Niyogi SK. Shigellosis. Journal of microbiology (Seoul, Korea). 2005; 43(2): 133-43.
18. Limmathurotsakul D, Wongratanacheewin S, Teerawattanasook N, Wongsuvan G, Chaisuksant S, Chetchotisakd P, et al. Increasing incidence of human melioidosis in northeast thailand. The American Journal of Tropical Medicine and Hygiene 2010; 82(6): 1113-7.
19. Oloomi M, Bouzari S, Ajdary S. Immune responses of mice immunized with active recombinant shiga toxin and its derivatives. Iran J Allergy Asthma Immunol. 2008;7(2):53-60.
20. Cruz-Migoni A, Hautbergue GM, Artymiuk PJ, Baker PJ, Bokori-Brown M, Chang C-T, et al. A Burkholderia pseudomallei toxin inhibits helicase activity of translation factor eIF4A. Science. 2011; 334(6057): 821-4.
21. Strockbine NA, Jackson M, Sung L, Holmes R, O'Brien AD. Cloning and sequencing of the genes for Shiga toxin from Shigella dysenteriae type 1. Journal of Bacteriology. 1988;170(3):1116-22.
22. Su G, Li F, Huang P, Rui X, Huang C. High level expression of Shiga toxin B subunit of Shigella dysenteriae serotype 1 in Escherichia coli. Chinese journal of biotechnology. 1992;9(1):49-55.
23. Kurohane K, Kobayashi C, Imai Y. Facilitated Production of Secretory IgA against Shiga Toxin B Subunits by Intranasal Application of Antigen‐Coated Polystyrene Microspheres. Microbiology and immunology. 2005;49(2):149-54.
24. Zhu C, Yu J, Yang Z, Davis K, Rios H, Wang B, et al. Protection against Shiga toxin-producing Escherichia coli infection by transcutaneous immunization with Shiga toxin subunit B. Clinical and Vaccine Immunology. 2008;15(2):359-66.
25. Johansson D, Kosovac E, Moharer J, Ljuslinder I, Brännström T, Johansson A, et al. Expression of verotoxin-1 receptor Gb3 in breast cancer tissue and verotoxin-1 signal transduction to apoptosis. BMC cancer. 2009;9(1):67.
26. Honari H, Minaei M. Cloning and Expression of Fusion Genes of Domain A-1 Protective Antigen of Bacillus Anthracis and Shigella Enterotoxin B Subunit (Stxb) In E. coil. SSU_Journals 2015; 22(6): 1702-11. [Persian]
27. Honari H, Baranvand M, Arefpour M, Hashemzadeh M, Pourhakak H, Minaei M, et al. Comparison of Antibody Titers against the Single, Mixed, Fused and Recombinant proteins, StxB, IpaD and StxB- IpaD. journal of ilam university of medical sciences. 2015;22(7):87-95. (Persian)
28. Engedal N, Skotland T, Torgersen ML, Sandvig K. Shiga toxin and its use in targeted cancer therapy and imaging. Microbial biotechnology. 2011;4(1):32-46.
29. Malina A, Cencic R, Pelletier J. Targeting translation dependence in cancer. Oncotarget. 2011;2(1-2):76-88.

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