Intein-Mediated Fusion Expression In Escherichia Coli And Antiviral Activity Of Recombinant Goose IFN-α

Interferon (interferon, IFN) is a glucoprotein which secreted by the cells with a broad-spectrum anti-viral, anti-cell division, immunomodulatory activity and in different ways affect cell metabolism, growth. Recently, people are more and more concerned about the effects of interferons on disease prevention and cure of poultry, because of the characters of non-medicine and non-toxic side effects. However, the research and application of GoIFN-αis behindhand relatively, and the expression of GoIFN-αin Ecoli. is pending further study.So far, E. coli is the most extensively used host for the production of recombinant proteins. However, most of the eukaryotic protein are tupically obtained as insoluble inclusion bodies that need solubilization and refolding. So the soluble expression of rGoIFN-αwith activity is more valuble for the production and application.In this study, we integrated the beneficial conditions of the soluble expression of recombinant protein in E. coli, then constructed a series of expression vectors. After contrastive analysis, we established the plan of soluble expression: we had connected the Ssp DnaB mini-intein (SDI) coding sequence with the GoIFN-αcDNA, and then merged NusA tag at the upstream of SDI-GoIFN-αsequence. Finally, the fragment of NusA-SDI-GoIFN-αwas digested ligated into the expression vector pColdⅢ, and we succeeded to constructing the intein-cold shock expression plasmid, named as pColdⅢ-SDI-GoIFN-α. The result showed that the fusion protein was expressed in the form of solubility. Overall, successful expression is of significance for the purification and production of GoIFN-α.