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International Journal of Molecular and Cellular Medicine، جلد ۵، شماره ۱، صفحات ۳۷-۴۸
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عنوان انگلیسی High Yield Overexpression, Refolding, Purification and Characterization of Pseudomonas aeruginosa Type B-Flagellin: An Improved Method Without Sonication
چکیده انگلیسی مقاله Pseudomonas aeruginosa as an opportunistic pathogen is a significant cause of acute and chronic infections in patients with compromised defenses. This bacterium is motile via a single polar flagellum made of polymerized flagellin subunits differentiated into two major serotypes: A and B. flagellin plays an important role as a virulence factor in the adhesion, colonization and invasion of P. aeruginosa into host epithelial cells. To develop a functional vaccine that can be used in practical application to prevent and treat infection, type B-flagellin was produced as recombinant protein. In this work, the fliC gene was introduced into a pET28a vector and expressed in Escherichia coli BL21 (DE3). The expressed recombinant protein was purified by a modified method without sonication using a HisTrap affinity column. The functional activities of produced flagellin were confirmed by ELISA, western blot analysis, motility inhibition assay and opsonophagocytosis test. The purification process of the type B-flagellin was lead to a high yield. The produced recombinant type B-flagellin showed high biological activity in all of these standard assays. In conclusions, this report provides the new protocol to efficiently obtain the type B-flagellin with high biological activity and immunogenicity. This immunogen can be introduced as an adjuvant or vaccine in the future study.
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نویسندگان مقاله سبحان فایزی | sobhan faezi
departments of mycobacteriology and pulmonary research, pasteur institute of iran, tehran, iran.

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

احمدرضا بهره مند | ahmad reza bahrmand
departments of mycobacteriology and pulmonary research, pasteur institute of iran, tehran, iran.

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

مهدی مهدوی | mehdi mahdavi
departments of immunology, pasteur institute of iran, tehran, iran.

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

سید داور سیادت | seyed davar siadat
departments of mycobacteriology and pulmonary research, pasteur institute of iran, tehran, iran.

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

ایرج نیکوکار | iraj nikokar
laboratory of microbiology and immunology of infectious diseases, paramedicine faculty, guilan university of medical sciences, guilan, iran.

سازمان اصلی تایید شده: دانشگاه علوم پزشکی گیلان (Guilan university of medical sciences)

سروش سرداری | soroush sardari
biotechnology research center, drug design and bioinformatics group, pasteur institute of iran, tehran, iran.

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

یان آلان holder | ian alan holder
departments of microbiology and surgery, college of medicine, university of cincinnati, and shriners burns institute, cincinnati, ohio, usa.
نشانی اینترنتی http://www.ijmcmed.org/browse.php?a_code=A-10-561-2&slc_lang=en&sid=en
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