A New Approach for Designing A Potentially Vaccine Candidate against Urinary Tract Infection by Using Protein Display on Lacto-bacillus Surface

• Main Authors: Jalil Fallah Mehrabadi, Gholamreza Goudarzi, Fatemeh Farahani Hatamabadi, Hedroosha Molla Agha Mirzaei, Sarah Najafi
• Format: Article
• Language: English
• Published: Tehran University of Medical Sciences 2013-07-01
• Series: Journal of Medical Bacteriology
• Online Access: http://jmb.tums.ac.ir/index.php/jmb/article/view/74
• ISSN: 2251-8649; 2322-2581

<p><strong><em>Background:</em></strong><strong> </strong>The prevalence of Urinary Tract Infection (UTI) is really high in the world. <em>Escherichia coli</em> is a major agent of UTI. One of the strategies for decreasing UTI infections is vaccine development. As the attachment is a really important stage in colonization and infection, at­tachment inhibition has an applied strategy. FimH protein is a major factor during bacterial colonization in urinary tract and could be used as a vaccine. Thus, it was considered in this research as a candidate antigen.</p> <p><strong><em>Methods: </em></strong>The sequences of <em>fimH</em> and <em>acmA</em> genes were used for designing a synthetic gene. It was cloned to pET23a expression vector and transformed to <em>E. coli</em> (DE3) Origami. To confirm the expression of recombinant protein, SDS-PAGE and western blotting methods were used. Subsequently, recombinant protein was purified. On the other hand, <em>Lactobacillus</em><em> reuteri</em> was cultured and mixed with FimH / AcmA recombinant protein. The rate of protein localization on lactobacillus surface was assessed using ELISA method.</p> <p><strong><em>Results:</em></strong> It was showed that the recombinant protein was expressed in <em>E. coli</em> (DE3) Origami and purified by affinity chromatography. Moreover, this protein could be localized on lactobacillus surface by 5 days.</p> <strong><em>Conclusion:</em></strong> In current study, a fusion recombinant protein was pre­pared and displayed on <em>L. reuteri</em> surface. This strain could be used for animal experiment as a competitor against Uropathogenic <em>E. coli</em> (UPEC). Using manipulated probiotics strains instead of antibiotic ther­apy could decrease the antibiotic consumption and reduce multi-drug resistant strains.<span style="font-family: Times New Roman; font-size: small;"><br /></span>