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

Authors

Ameneh Koochaki, Shahid Beheshti University of Medical Sciences
Mojgan Bandehpour, Shahid Beheshti University of Medical SciencesFollow
Sam Massahi, Shahid Beheshti University of Medical Sciences
Mahrou Sadri, University of Nebraska-Lincoln
Bahram Kazemi, Shahid Beheshti University of Medical Sciences

Date of this Version

2015

Citation

Koochaki A, Bandehpour M, Massahi S, Sadri M, Kazemi B. Cloning and Expression of EBI3 and p28 Subunits of Human Interleukin 27 in E. coli. Novel Biomed. 2015;3(3):140-3.

Comments

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

Abstract

Background: 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.

Materials and Methods: 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.

Results: 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.

Conclusion: In this research p28 and EBI3 proteins were produced successfully.