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Volume 7, Issue 1 (Feb 2019)                   Res Mol Med (RMM) 2019, 7(1): 0-0 | Back to browse issues page

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Abdi A, Hosseinpour M, Mashayekhi K, Mousavi M J, Badiee Kheirabadi S E, Sankian M. Optimization of Cloning Conditions for high-level Production of Recombinant Mouse Interleukin-2 in Escherichia coli. Res Mol Med (RMM). 2019; 7 (1)
URL: http://rmm.mazums.ac.ir/article-1-309-en.html
Abstract:   (169 Views)
Backgrounds and objectives: Interleukin 2 (IL-2) secreted by activated CD4+ T cells  has been known as a major mediator in both  adaptive and native immune system  due to a board range of effects on  different cells in the immunity system (1-6).
Methods: cDNA synthesis was performed using gene- specific primers designed by Gene Runner software after RNA extraction of mouse splenocytes. PCR product purification   carried out after IL-2 coding sequence amplification and was ligated into the pET-21b (+) vector and transformed into competent TOP10 E.Coli. Expression and purification of recombinant mouse IL-2 were done using IPTG inducer and metal affinity chromatography respectively.
Results: DNA sequencing confirmed the accuracy of the insertion process. 23 KDa exogenous protein was observed on the SDS-PAGE. Specificity and   concentration of produced mouse recombinant IL-2 protein were confirmed by western blotting and BCA methods.
Conclusion: Recombinant IL-2 has produced in BL21 (DE3) pET-21b (+) expression system at 24°C in the soluble form.
 
     
Type of Study: Research | Subject: Immunology
Received: 2019/05/24 | Accepted: 2019/07/7

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