Abstract
Interferon alpha 2b (IFNα-2b) is an important cytokine and used for antiviral and anticancer treatment. The low cost production of IFNα-2b with high biological activity is necessary to provide the interferon therapy to the hepatitis patients in Pakistan. In the present study, human interferon alpha 2b (hIFNα-2b) gene from a healthy person was cloned and overexpressed in E. coli BL21(DE3). The molecular weight of the expressed hIFNα-2b is 19 kDa. The over expressed recombinant hIFNα-2b was checked by ELISA using antibodies raised against commercially available hIFNα-2b. The biocomputational analysis of recombinant hIFNα-2b gene showed the 99.9% nucleotide sequence and 100% deduced amino acid sequence homology with reported sequences of IFNα-2b. The predicted 3D-structure showed mainly five α-helices, one 310 helix and two disulfide bonds at Cys1-Cys98 and Cys129-Cys138. The amino acid sequence alignment indicated that the disulfide linkage position is conserved in all IFNα family members. On the basis of sequence homology among interferon alpha family, new potent variants of hIFNα-2b with enhance efficacy can be produced. Indigenous production of IFNα-2b from gene of local population will reduce the cost and increase tolerability of interferon therapy.
Highlights
Interferons (IFNs) are multigene family of inducible cytokines which are produced in response to stimulation by certain viruses, bacteria, antigens and mitogens
Cloning and characterization of hIFNα-2b gene The gene fragment of 567 bp encoding human IFNα-2b was amplified by RT-PCR from leukocytes isolated from peripheral blood of healthy person
Hepatitis C is an emerging health problem worldwide and it has been estimated that Hepatitis C virus (HCV) has infected approximately 17 million people in Pakistan (Akbar et al 2009)
Summary
Interferons (IFNs) are multigene family of inducible cytokines which are produced in response to stimulation by certain viruses, bacteria, antigens and mitogens. Type I IFNs are known as viral IFNs, which include IFN-α, IFN-β, IFN-ω, IFN-ε, IFN-ν and IFN-κ. These are considered as primary line of defense of the host immune system against infectious agents and tumour progression (Salunkhe et al 2010). Type II IFN, known as immune IFN, which includes IFN- γ and induced by mitogenic or antigenic stimuli (Pestka 2007). The type I IFNs gene cluster is located on the short arm of Homo sapiens chromosome 9 (9p21) (Diaz et al 1994). They consist of 26 genes including 13 IFN-α genes, 1 IFNβ gene, 1 IFN-ω gene and 11 IFN pseudogenes (Roberts et al 1998).
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