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Cloning and Expression of EBI3 and p28 Subunits of Human Interleukin 27 in E. coli

Ameneh Koochaki, Mojgan Bandehpour, Sam Massahi, Bahram Kazemi
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Abstract

Background: Interleukin-27 is a heterodimeric cytokine belonging to IL-12 and IL-23 families, secreted  by Antigen Presenting Cells(APCs) .The IL-27 is composed of 2 subunits: Epstein-Barr virus induced gene 3 (EBI3) and p28. IL-27 is an anti-inflammatory cytokine which has an inhibitory effect on Th17 population and suppress the IL-17 expression. It is suggested that IL-27 could be a potent drug candidate for treating auto immune diseases.

Methods: The IL-27 subunit genes were constructed into plasmid vectors, they sub-cloned into pETDeut-1an expression vector in restriction sites of BamHI, SacI and NotI. Subsequently the induction was done by IPTG and the recombinant proteins were confirmed by SDS-PAGE and Western Blot analysis using anti His-tag antibody.

Results: The 28kDa and 25kDa protein bands were observed on SDS-PAGE and finally confirmed by Western blot technique.


Keywords

Cytokine, Recombinant protein, Autoimmune disease

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DOI: https://doi.org/10.22037/nbm.v3i3.7304