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Iranian Journal of Medical Sciences، جلد ۲۸، شماره ۳، صفحات ۱۳۱-۰
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عنوان انگلیسی Overexpression of Recombinant Human Granulocyte Colony-Stimulating Factor in E. coli
چکیده انگلیسی مقاله Bakground: Granulocyte colony-stimulating factor (G-CSF) is a cytokine that stimulates hematopoiesis and induces proliferation and differentiation of granulocyte progenitor cells as well as production of bone marrow neutrophilic granulocyte colonies. Nowadays, human recombinant G-CSF(hr G-CSF) is used for the treatment of chemotherapy- and radiotherapy-induced neutropenia, and also in patients with bone marrow transplantation. Methods: A cDNA of human G-CSF (hG-CSF) was synthesized by PCR from recombinant cloning vector, with two altered nucleotides for increasing mRNA stability and overexpression, then inserted into a pET expression vector under the control of T7 promoter and cloned in E. coli strain BL21 (DE3). Results: After culture and induction of recombinant E. coli with IPTG, we achieved a high level expression of the hG-CSF, where it represented approximately 35% of the total protein as determined by SDS-PAGE and confirmed by western blotting with polyclonal and monoclonal hG-CSF antibodies. Conclusion: rhG-CSF was produced in a significantly high quantity with a yield of 35% of total protein as determined by SDS-PAGE. Since it is easily obtained by simple purification steps, it may be cost-effective, even on an industrial scale.
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نویسندگان مقاله m j فلاح | m j fallah
department of genetic engineering, research center for science and biotechnology, tehran, iran


b اکبری | b akbari
department of genetic engineering, research center for science and biotechnology, tehran, iran


a r سعیدی نیا | a r saeedinia
department of genetic engineering, research center for science and biotechnology, tehran, iran


کریمی | m karimi
department of biotechnology, pasteur institute of iran, tehran, iran

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

واعظ | m vaez
department of genetic engineering, research center for science and biotechnology, tehran, iran


m زین الدینی | m zeinoddini
department of genetic engineering, research center for science and biotechnology, tehran, iran


مجید سلیمانی | m soleimani
department of hematology, tarbiat modarres university, school of medicine,tehran, iran

سازمان اصلی تایید شده: دانشگاه تربیت مدرس (Tarbiat modares university)

n مقصودی | n maghsoudi
department of genetic engineering, research center for science and biotechnology, tehran, iran
نشانی اینترنتی http://ijms.sums.ac.ir/index.php/IJMS/article/view/2467
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زبان مقاله منتشر شده en
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