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:: دوره 17، شماره 2 - ( 12-1393 ) ::
برگشت به فهرست نشریات جلد 17 شماره 2 صفحات 47-51
Expression Optimizing and Purification of Recombinant Human Leukemia Inhibitory Factor Produced in E. coli Strain BL21
چکیده:   (6340 مشاهده)
Background : Leukemia inhibitory factor (LIF) is a glycoprotein, categorized as a subfamily of interleukin 6 cytokines which is known in many mammals. A pluripotent cytokine with a wide biological function range has numerous effects on target cells. The LIF regulates neuron survival, hematopoiesis and seen in LIF-/- knockout mice affects blastocyst implantation, also acts as pre-inflammatory cytokine, and regulates immune response. Further, it is able to maintain stem cells poly potency. The main object of present work was expression, optimizing, and purification of recombinant human leukemia inhibitory factor (rhLIF).
Materials and Methods : In this experimental study, Pet28 (+) carrying the LIF gene and kanamycin resistance marker was cloned in E. coli strain BL21. The induction was optimized by altering 3 factors including the temperature, the induction time, and the concentration of the Isopropyl β-D-1-thiogalactopyranoside (IPTG) as inducer. The purification of the recombinant human LIF (rhLIF) was done by single step affinity chromatography. After the purification, method accuracy was proved by Sodium dodecyl sulfate (SDS) -PAGE electrophoresis and Western blotting.
Results : Optimizing of the expression was reached by changing various parameters, and purification has been done successful.
Conclusion : rhLIF undergoes modification by glycosylation to get its full functionality. The produced rhLIF in prokaryotic host in this work is lacking of glycosylation. However, its proper function should be evaluated in further studies.
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نوع مطالعه: پژوهشي | موضوع مقاله: ژنتیک وبیو لوژی مولکولی
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Kahroba H, Galehdari H, Shafeei M, Khatami S R, Khodadadi A. Expression Optimizing and Purification of Recombinant Human Leukemia Inhibitory Factor Produced in E. coli Strain BL21. ZJRMS. 2015; 17 (2) :47-51
URL: http://zjrms.ir/article-1-3084-fa.html
Expression Optimizing and Purification of Recombinant Human Leukemia Inhibitory Factor Produced in E. coli Strain BL21. Zahedan Journal of Research in Medical Sciences. 1393; 17 (2) :47-51
برگشت به فهرست نشریات دوره 17، شماره 2 - ( 12-1393 )
Zahedan Journal of Research in Medical Sciences
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