Cloning and expression of EBI3 and p28 subunits of human interleukin 27 in E. coli

ABSTRACT

Interleukin [IL]-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. The EBI3 and p28 subunits of human interleukin 27 were constructed into plasmid vectors; they sub-cloned into pETDuet-1 expression vector in restriction sites of BamHI, SacI and NotI. Subsequently the induction was carried out by 1mM IPTG and the recombinant proteins were confirmed by SDS-PAGE and Western Blot analysis using anti His-tag antibody. The EBI3 and p28 subunits of human interleukin 27were cloned into plasmid vectors. The 28 and 25kDa protein bands were observed on SDS-PAGE and finally confirmed by Western blot technique. In this research p28 and EBI3 proteins were produced successfully