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cloning, expression and purification of the factor h binding protein and its interaction with factor h
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نویسنده
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yarian fatemeh ,bandehpour mojgan ,seyed negar ,kazemi bahram
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منبع
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iranian journal of microbiology - 2016 - دوره : 8 - شماره : 1 - صفحه:29 -35
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چکیده
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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 fhbp 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.
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کلیدواژه
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n. meningitidis ,fhbp ,cloning ,fh protein ,far-western blotting
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آدرس
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department of biotechnology, school of medicine, shahid beheshti university of medical sciences, tehran ,iran., Iran, Islamic Republic of, 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., Iran, Islamic Republic of, department of immunotherapy and leishmania vaccine research, pasteur institute of iran, tehran, iran., Iran, Islamic Republic of, 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 ofmedical sciences, tehran, iranand cellular and molecular biology research center, shahid beheshti university of medical sciences, tehran, iran., Iran, Islamic Republic of
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پست الکترونیکی
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bandehpour@gmail.com
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Authors
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