Cloning and Expression of EBI3 and p28 Subunits of Human Interleukin 27 in E. coli
Novelty in Biomedicine,
Vol. 3 No. 3 (2015),
22 July 2015
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.
- Recombinant protein
- Autoimmune disease
How to Cite
Jankowski M, Kopiński P, Goc A. Interleukin-27: Biological Properties and Clinical Application. Arch ImmunolTher Exp. 2010;58:417–25.
Diveu C, McGeachy M, Boniface K, et al. IL-27 blocks RORc expression to inhibit lineage commitment of Th17 Cells. J Immunol. 2009;182:5748.
Pflanz S, Timans JC, Cheung J, et al. IL-27, a heterodimeric cytokine composed of EBI3 and p28 protein, induces proliferation of naive CD4+ T cells. Immunity. 2002;16:779–90.
Yuan X, Hu J, Yoshimoto T et al. Bone marrow derived neural stem-like cells expressing IL-27 exhibit antitumor activity in intracranial gliomas. MolTher.2005;11:S268.
Shimizu M, Shimamura M, Owaki T, et al. Antiangiogenic and antitumor activities of IL-27. J Immunol. 2006;176:7317–24.
Batten M, Li J, Yi S, et al. Interleukin 27 limits autoimmune encephalomyelitis by suppressing the development of interleukin 17-producing T cells. Nat Immunol. 2006;7:929–36.
Fitzgerald DC, Ciric B, Touil T, et al. Suppressive effect of IL-27 on encephalitogenic Th17 cells and the effector phase of experimental autoimmune encephalomyelitis. J Immunol. 2007;179:3268–3275.
Furukawa M, Takaishi H, Takito J et al.IL-27 abrogates receptor activator of NF-kappaB ligand-mediated osteoclastogenesis of human granulocyte-macrophage colony-forming unit cells through STAT1-dependent inhibition of c-Fos. J Immunol. 2009;183:2397–406.
Fakruddin JM, Lempicki RA, Gorelick RJ, et al. Noninfectious papilloma virus-like particles inhibit HIV-1 replication: implications for immune control of HIV-1 infection by IL-27. Blood. 2007;109:1841–9.
Bender H, Wiesinger M, Nordhoff C, et al. Interleukin-27 displays interferon-gamma-like functions in human hepatoma cells and hepatocytes. Hepatol. 2009;50:585–91.
Greenwell-Wild T, Vazquez N, Jin W, et al. IL-27 inhibition of HIV-1 involves an intermediate induction of type I interferon. Blood. 2009;114:1864–74.
Lucas S, Ghilardi N, Li J, et al. IL-27 regulates IL-12 responsiveness of naive CD4+ T cells through Stat1-dependent and -independent mechanisms. Proc Natl Acad Sci USA. 2003;100:15047–52.
Marcel B. The biology and therapeutic potential of interleukin 27. J Mol Med. 2007;85:661–72.
Hiroki Y, Mako N, Yoshiyuki M.Interleukin 27: a double-edged sword for offense and defense. Leukocyte Biol. 2009;86:1295–303.
Bandehpour M, Khodabandeh M, Kazemi B. Cloning and Expression of Hepatitis B virus Surface Antigen. Hepat Month. 2008;8(1):17-21.
Hu P, Peng M, Chen M, Hu H, Tang L, Tang K, Ren H. Molecular cloning, expression and characterization of an interleukin27 p28 homolog in pig (Susscrofa). Veterinary Immunol and Immunopath. 2009;130:262–7.
Lotfipour F, Hassanpour R, Farajnia S. Cloning and Expression of human IL-11 in E. coli. Pharma Sci. 2010;15: 353-9.
Tan H, Dan G, Gong H, Cao L. Purification and characterization of recombinant truncated human interleukin-11 expressed as fusion protein in Escherichia coli. Biotechnology Letters. 2005;27:905-10.
- Abstract Viewed: 616 times
- PDF Downloaded: 417 times