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Journal of Paramedical Sciences، جلد ۳، شماره ۲، صفحات ۰-۰
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| عنوان فارسی |
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| چکیده فارسی مقاله |
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| عنوان انگلیسی |
HPV16 E7-CT (gp96) fusion protein: Molecular cloning, expression and purification of a recombinant 6xHis-tagged protein in E. coli |
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| چکیده انگلیسی مقاله |
The development of a therapeutic vaccine against human papillomavirus (HPV) is important for the control of cervical cancer. E7 is the major transforming protein produced in cervical cancers, and therefore represents potential tumor-specific antigen that could be the target of immunotherapy for cervical cancer. Among different vaccine strategies, protein-based vaccines are capable of generating CD8+ T cell responses in vaccinated animals and humans. Recently, development of novel strategies that enhance protein vaccine potency is important for generation of effective cancer vaccines and immunotherapies. Heat shock proteins (HSPs) including Gp96 have been shown to act as potent immuno-adjuvant to enhance antigen-specific tumor immunity. Therefore, the HSP-based protein vaccines can be administered by fusing antigens to HSPs, in vitro . It has been known that the HSP fragments (e.g., N-/or C-terminal regions) as mini-chaperones are better choice for immunization. The most straightforward method to produce large amounts of recombinant protein suitable for a vaccine is to clone the gene into a prokaryotic expression vector and produce the protein in Escherichia coli . In current study, we describe cloning of the HPV16 E7 gene linked to C-terminal fragment of gp96, identification and purification of the resultant E7-CT (gp96) fusion protein for next usage as a potential vaccine candidate protein against HPV in a pre-clinical trial. The recombinant E7-CT (gp96) migrated as a 51 kDa protein in SDS-PAGE. In Western blot experiment, the existence of a 51 kDa band for rE7-CT (gp96) was confirmed by rabbit anti-His as well as mouse anti-HPV16 E7 monoclonal antibodies. The protein of interest was both in the insoluble and the soluble fraction; therefore, purification was performed under denaturating and native conditions by affinity chromatography on Ni-NTA resin using 6xHis-tag. |
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| کلیدواژه‌های انگلیسی مقاله |
HPV,E7,Gp96,cervical cancer,protein expression,E.coli |
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| نویسندگان مقاله |
امین دایمی | amin daemi
molecular immunology and vaccine research lab., pasteur institute of iran, tehran
سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)
سحر حسین زاده | sahar hosseinzadeh
islamic azad university of pharmaceutical science branch, tehran
سازمان اصلی تایید شده: دانشگاه آزاد اسلامی علوم و تحقیقات (Islamic azad university science and research branch)
سیما رفعتی | sima rafati
molecular immunology and vaccine research lab., pasteur institute of iran, tehran
سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)
فرناز زاهدی فرد | farnaz zahedifard
molecular immunology and vaccine research lab., pasteur institute of iran, tehran
سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)
معصومه رجبی بذل | masoumeh rajabi bazl
department of clinical biochemistry, shahid beheshti university of medical sciences
سازمان اصلی تایید شده: دانشگاه علوم پزشکی شهید بهشتی (Shahid beheshti university of medical sciences)
فاطمه doustdari | fatemeh doustdari
molecular immunology and vaccine research lab., pasteur institute of iran, tehran
سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)
مهرداد هاشمی | mehrdad hashemi
department of genetics, islamic azad university, tehran medical branch
سازمان اصلی تایید شده: دانشگاه آزاد اسلامی علوم و تحقیقات (Islamic azad university science and research branch)
الناز agi | elnaz agi
molecular immunology and vaccine research lab., pasteur institute , tehran
سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)
اعظم بوالحسنی | azam bolhassani
molecular immunology and vaccine research lab., pasteur institute of iran, tehran,
سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)
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| نشانی اینترنتی |
http://journals.sbmu.ac.ir/jps/article/view/3271 |
| فایل مقاله |
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| کد مقاله (doi) |
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| زبان مقاله منتشر شده |
en |
| موضوعات مقاله منتشر شده |
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| نوع مقاله منتشر شده |
Research/Original Articles |
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