Cloning, Expression And Characterization Of New Type Recombinant Human Interferon-α

Iiiterferon (IFN) was first discovered as a substance that can induce an antiviral state in cells. Since then, many types of IFNs have been isolated and classified according to cell surface receptors, acid stability, primary sequence, and chromosomal localization. IFNs are divided into two major groups, type I and type II. The type I IFN family is comprised of 1FN-α and IFN-β In. humans, at least 14 nonallelic genes encode structurally different forms of IFN-α, including one pseudogene and two genes that encode the same protein. Members of the type I IFN family have homology to each other, bind to the same cell surface receptors and have overlapping functions. Their broad range of biological properties has suggested many clinical applications in cancer and virology diseases. New recombinant human Interferon alpha (rIFN α ) which is absent in nature was designed as 166 amino acids with arginine at 122 site. According to the product of American Amgen Company, the rIFNa has the potential antiviral activity. The express of the gene of rIFNα in E. coli system and biological activities were investigated.Clone of rIFNα gene: The sequence of complete DNA was designed to separate as eight fragments with similar length, including four right chain fragments (F1-F4) and four reverse chain fragments (R1-R4) which were automatically synthesized. Those fragments were complementation with designed one, respectively. The gene was amplified by PCR and then was subcloned into pUC18 vector to construct recombinant expression plasmid. The sequence was analyzed and was same as that designed target DNA.Expression of rIFNα in E. coli : IFNα gene was inserted into the expression...