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Iranian Journal of Microbiology، جلد ۸، شماره ۱، صفحات ۲۹-۳۵
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عنوان انگلیسی Cloning, expression and purification of the factor H binding protein and its interaction with factor H
چکیده انگلیسی مقاله Background and Objective: Neisseria meningitidis is a leading cause of meningitis and sepsis worldwide. The factor H binding protein (fHBP) is a key virulence factor of Neisseria meningitidis that is able to selectively bind to human factor H, the key regulator of the alternative complement pathway, which it has important implications for meningococcal pathogene- sis and vaccine design. The aims of present research were cloning, expression, purification of fHbp and confirmation of the interaction between serum factor H (fH) and produced factor H binding protein. Materials and Methods: A 820 base pairs fh bp gene fragment was amplified by PCR and cloned into expression vector pE- T28a (+) in Bam HI and SalI restriction enzymes sites. Recombinant DNA was expressed in BL21 (DE3) cell. fHBP protein was purified by Ni-NTA agarose resin. Coupling of recombinant protein into CNBr activated Sepharose 4B resin was carried out for application in serum fH protein purification. (fH-fHBP) interaction was confirmed by SDS-PAGE and far-western blotting. Results and Conclusions: SDS-PAGE results showed a 35 kDa protein band. 150 kDa fH protein was purified by designed Sepharose 4B resin. Far-western blotting confirmed (fH-fHBP) interaction and proper folding of factor H binding protein.
کلیدواژه‌های انگلیسی مقاله N. meningitidis,fHbp,cloning,fH protein,far-western blotting
نویسندگان مقاله فاطمه یاریان | fatemeh yarian
department of biotechnology, school of medicine, shahid beheshti university of medical sciences, tehran ,iran.

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شهید بهشتی (Shahid beheshti university of medical sciences)

مژگان بنده پور | mojgan bandehpour
department of biotechnology, school of advanced technologies in edicine, shahid beheshti university of medical sciences, tehran, iran andcellular and molecular biology research center, shahid beheshti university of medical sciences, tehran, iran.

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شهید بهشتی (Shahid beheshti university of medical sciences)

نگار سید | negar seyed
department of immunotherapy and leishmania vaccine research, pasteur institute of iran, tehran, iran.

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

بهرام کاظمی | bahram kazemi
department of biotechnology, school of medicine, shahid beheshti university of medical sciences, tehran, iran and department of biotechnology, school of advanced technologies in medicine, shahid beheshti university of medical sciences, tehran, iran and cellular and molecular biology research center, shahid beheshti university of medical sciences, tehran, iran.

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شهید بهشتی (Shahid beheshti university of medical sciences)

نشانی اینترنتی http://ijm.tums.ac.ir/index.php/ijm/article/view/724
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