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Iranian Journal of Biotechnology، جلد ۱۱، شماره ۱، صفحات ۴۷-۵۳
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عنوان انگلیسی Codon Optimization, Cloning and Expression of the Human Leukemia Inhibitory Factor (hLIF) in E. coli
چکیده انگلیسی مقاله Background: Leukemia inhibitor factor (LIF) is a very important pleiotropic cytokine which belongs to interleukin-6 (IL-6) family. LIF exerts multiple effects on different types of cells and tissues with numerous regulatory effects in vivo and in vitro. It is a lymphoid factor, which performs a number of activities including cholinergic neuron differentia­tion, control of stem cell pluripotency, bone and fat metabolism, and is important for embryo implantation and promoting megakaryocytes production in vivo. Human LIF is a potential therapeutic candidate for some diseases such as multiplesclerosis (MS). Objectives: Because of aforementioned applications of the LIF protein in biological sys­tems, the LIF gene has been cloned in various species. In this study a useful novel method was used to clone an optimized LIF sequence. Materials and Methods: In this study, the optimized cDNA form of human leukemia inhibitory factorwas cloned into the pET-28a (+) expression vector under control of T7lacpromoter using BamHI and XbaI restriction enzymes. The recombinant vector was transformed into Escherichia coli strain BL21 (DE3). The cloned hLIF cDNA was expressed as a fusion protein with His-tag. Cloning of hLIF cDNA was confirmed by digestion and DNA sequencing. Appropriate expression of recombinant hLIF was examined by SDS-PAGE af­ter induction with isopropylthio-β-galactoside (IPTG). Results: The results confirmed the expression of the 19.7 kDa rhLIF protein in the bacte­rial expression system. Conclusions: We could show that codon optimization might increase the production of recombinant hLIF in the E. coli. This result is useful in similar cases, in which the level of expressed gene is critical.
کلیدواژه‌های انگلیسی مقاله Escherichia coli,Inhibitor Factor
نویسندگان مقاله پریچهر دارابی |
department of genetics, shahid chamran university, ahvaz, ir iran
سازمان اصلی تایید شده: دانشگاه شهید چمران (Shahid chamran university)

حمید گله داری |
department of genetics, shahid chamran university, ahvaz, ir iran
سازمان اصلی تایید شده: دانشگاه شهید چمران (Shahid chamran university)

سعیدرضا خاتمی | saeed reza
department of genetics, shahid chamran university, ahvaz, ir iran
سازمان اصلی تایید شده: دانشگاه شهید چمران (Shahid chamran university)

ناهید شهبازیان |
department of gynecology, ahvaz jundishapur university of medical school, ahvaz, ir iran

محمد شافعی |
department of genetics, shahid chamran university, ahvaz, ir iran
سازمان اصلی تایید شده: دانشگاه شهید چمران (Shahid chamran university)

امیر جلالی |
research center of toxicology, ahvaz jundishapur university of medical school, ahvaz, ir iran

علی خدادادی |
cancer research center, ahvaz jundishapur university of medical school, ahvaz, ir iran

نشانی اینترنتی http://www.ijbiotech.com/article_7195_ee0ae53881bbaa801b146bc83f5edd2c.pdf
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