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Avicenna Journal of Medical Biochemistry، جلد ۲، شماره ۱، صفحات ۰-۰
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عنوان فارسی |
Cloning and Expression of Human Keratinocyte Growth Factor in Escherichia coli for Recombinant Drug Production |
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چکیده فارسی مقاله |
Results Cloning was confirmed by PCR and restriction digestion. RT-PCR and SDS-PAGE represented expression of KGF in E. coli. The optimized expression was achieved 16 hours after induction with 0.3 mM IPTG at 37°C in luria broth (LB) containing kanamycin. The 18 kDa protein was confirmed by western blotting, using anti-His antibodies. Conclusions The result of the present study indicated that E. coli expression system was suitable for overexpression of recombinant human KGF and the expressed protein can be considered as a homemade product. Materials and Methods KGF gene was amplified by PCR and cloned into the expression vector pET28a(+). The recombinant vectors were transformed into E. coli BL21(DE3) as expression host and expression of the desired protein was induced by IPTG. The expression was evaluated at RNA and protein levels by reverse transcriptase PCR (RT-PCR) and SDS-PAGE analyses, respectively and the expressed protein was confirmed through western blotting. Objectives The aim of this study was to express the recombinant form of human keratinocyte growth factor in Escherichia coli. Background Keratinocyte growth factor (KGF) is a member of fibroblast growth factor (FGF) family which induces proliferation and differentiation in a wide variety of epithelial tissues. KGF plays an important role in protection, repair of various types of epithelial cells, and re-epithelialization of wounds. Therefore, in patients with hematologic malignancies receiving high doses of chemotherapy and radiotherapy, treatment with KGF decreases the incidence and duration of severe oral mucositis. |
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کلیدواژههای فارسی مقاله |
Cloning،Recombinant Protein،Keratinocyte Growth Factor |
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عنوان انگلیسی |
Cloning and Expression of Human Keratinocyte Growth Factor in Escherichia coli for Recombinant Drug Production |
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چکیده انگلیسی مقاله |
Results Cloning was confirmed by PCR and restriction digestion. RT-PCR and SDS-PAGE represented expression of KGF in E. coli. The optimized expression was achieved 16 hours after induction with 0.3 mM IPTG at 37°C in luria broth (LB) containing kanamycin. The 18 kDa protein was confirmed by western blotting, using anti-His antibodies. Conclusions The result of the present study indicated that E. coli expression system was suitable for overexpression of recombinant human KGF and the expressed protein can be considered as a homemade product. Materials and Methods KGF gene was amplified by PCR and cloned into the expression vector pET28a(+). The recombinant vectors were transformed into E. coli BL21(DE3) as expression host and expression of the desired protein was induced by IPTG. The expression was evaluated at RNA and protein levels by reverse transcriptase PCR (RT-PCR) and SDS-PAGE analyses, respectively and the expressed protein was confirmed through western blotting. Objectives The aim of this study was to express the recombinant form of human keratinocyte growth factor in Escherichia coli. Background Keratinocyte growth factor (KGF) is a member of fibroblast growth factor (FGF) family which induces proliferation and differentiation in a wide variety of epithelial tissues. KGF plays an important role in protection, repair of various types of epithelial cells, and re-epithelialization of wounds. Therefore, in patients with hematologic malignancies receiving high doses of chemotherapy and radiotherapy, treatment with KGF decreases the incidence and duration of severe oral mucositis. |
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کلیدواژههای انگلیسی مقاله |
Cloning,Recombinant Protein,Keratinocyte Growth Factor |
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نویسندگان مقاله |
فاطمه ابراهیم زاده | fatemeh ebrahimzadeh research center for molecular medicine, department of molecular medicine and genetics, hamadan university of medical sciences, hamadan, ir iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی همدان (Hamadan university of medical sciences)
یگانه طالب خان | yeganeh talebkhan department of biotechnology, biotechnology research center, pasteur institute of iran, tehran, ir iran
سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)
حسن میرزاحسینی | hassan mirzahoseini department of biotechnology, biotechnology research center, pasteur institute of iran, tehran, ir iran
سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)
قاسم براتی | ghasem barati research center for molecular medicine, department of molecular medicine and genetics, hamadan university of medical sciences, hamadan, ir iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی همدان (Hamadan university of medical sciences)
مسعود سعیدی جم | massoud saidijam research center for molecular medicine, department of molecular medicine and genetics, hamadan university of medical sciences, hamadan, ir iran; research center for molecular medicine, department of molecular medicine and genetics, school of medicine, hamadan university of medical sciences, hamadan, ir iran. tel 98-9121324616, fax 98-8138380464
سازمان اصلی تایید شده: دانشگاه علوم پزشکی همدان (Hamadan university of medical sciences)
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نشانی اینترنتی |
http://www.avicennajb.com/index.php?page=article&article_id=19002 |
فایل مقاله |
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کد مقاله (doi) |
10.17795/ajmb-19002 |
زبان مقاله منتشر شده |
fa |
موضوعات مقاله منتشر شده |
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نوع مقاله منتشر شده |
research-article |
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