en Medical Biotechnology Medical Biotechnology Original Article Original Article <div style="text-align: justify;">An increasing number of studies have demonstrated the effectiveness of flagellin (FliC) as an adjuvant. In this study, FliC(s) from <em>Salmonella enterica </em>serovar Typhimurium, <em>Pseudomonas aeruginosa</em>, and <em>Escherichia coli</em> were compared. <em>In silico</em> studies were designed to determine physicochemical parameters of FliC from different bacteria. The <em>fliC </em>gene corresponding to best model was amplified, cloned and expressed into pET28a vector. Bioactivity of the protein was determined by measurement of the interleukin 8 in cell culture after interaction of FliC with TLR-5. According to <em>in silico</em> studies, FliC of <em>S. typhimorium</em> has a higher quality of physiochemical properties, and more affinity to TLR-5 than the other FliCs. SDS-PAGE showed an approximately 54 kDa band, and Western blot confirmed the presence of recombinant protein. IL-8 was expressed significantly in cell culture under <em>S. typhimorium</em> FliC induction. Considering side effects, and costs of conventional adjuvants, FliC can be introduced as a potent natural adjuvant in future studies. <em>In silico</em> studies can save time and cost, and screen potent proteins.</div> Salmonella typhimurium, FliC, adjuvant 82 88 http://ibbj.org/browse.php?a_code=A-10-272-1&slc_lang=en&sid=1 Zakaria Bameri zakariabameri@yahoo.com 10031947532846003225 10031947532846003225 No Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran Mohammad Reza Asadi Karam zakariabameri@gmail.com 10031947532846003226 10031947532846003226 No Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran Mehri Habibi stud.z_bameri@pasteur.ac.ir 10031947532846003227 10031947532846003227 No Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran Mana Oloomi manaoloomi@yahoo.com 10031947532846003228 10031947532846003228 No Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran Parastoo Ehsani p_ehsani@yahoo.com 10031947532846003229 10031947532846003229 No Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran Saeid Bouzari saeidbouzarii@yahoo.com 10031947532846003230 10031947532846003230 Yes Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran