Tarbiat Modares University Pathobiology Research 2538-3000 23 4 2020 06 01 Design of vaccine DNA containing genes encoding sicl Escherichia coli antigens Enterotoxigenic (ETEC) and Escherichia coli enterohemorrhagic (EHEC) طراحی DNA واکسن حاوی ژن‌های کدکننده sicl آنتی‌ژن‌های باکتری اشرشیاکلی انتروتوکسیژنیک(ETEC) و اشریشیاکلی انتروهموراژیک(EHEC) 1 8 19386 EN Fatemeh Jafari Parsa Department of Genetics, Faculty of Basic Sciences, Islamic Azad University, Research Sciences Branch, Tehran, Iran. Shohre Zare Department of Genetics and Biotechnology, Pishva Faculty of Biological Sciences, Islamic Azad University, Varamin Branch, Iran. Seyed Ali Mirhosseini Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran- Iran. Jafar Amani Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Vanak Sq. Molasadra St. Tehran- Iran. Journal Article <strong>Introduction:</strong> Enterotoxigenic <em>Escherichia coli</em> (ETEC) and Enterohemorrhagic <em>Escherichia coli</em> O157:H7 , the most common strains, are causes diarrhea that can kill hundreds of thousands of children annually. The development of an effective combination vaccine for enterohemorrhagic <em>Escherichia coli</em> and Enterotoxigenic <em>Escherichia coli</em> is very important.<span dir="RTL"></span><br> <strong>Materials and Methods:</strong> In this study, the <em>sicl</em> gene was amplified and Subcloned as a DNA vaccine. The sequence of antigen encoding genes was evaluated from the genebank and their epitopes were evaluated to design of primer for the synthetic chimeric gene and was amplified by PCR. Subcloning of a multipartal chimeric gene in eukaryotic expression vector was performed to make a DNA vaccine and finally the protein was purified by nickel chromatography and evaluated by Western blotting.<span dir="RTL"></span><br> <strong>Results:</strong> The immunoblotting results of the expression of SICL chimeric protein indicated the presence of a 76 kDa band in the form of insoluble particles after 12 hours induction. Purification of the recombinant protein using His tag sequencee and confirmation of the purified protein with the recombinant protein specific antibody demonstrated the accuracy of the protein expression.<span dir="RTL"></span><br> <strong>Discussion & Conclusion:</strong> Protein expression, purification and verify by western blotting showed that this recombinant chimeric protein (SICL) can be expressed in eukaryotic host.<span dir="RTL"></span><span dir="RTL"></span> https://mjms.modares.ac.ir/article_19386_b62d8976329e9c1c304baa4427f8e0a2.pdf