Monday, July 27, 2009
P32

OVER EXPRESSION OF BIOLOGICALLY ACTIVE INTERFERON BETA USING SYNTHETIC GENE IN E. coli    OVER EXPRESSION OF BIOLOGICALLY ACTIVE INTERFERON BETA USING SYNTHETIC GENE IN E. coli    OVER EXPRESSION OF BIOLOGICALLY ACTIVE IN

Maryam Ghane1, Bagher Yakhchali2, and Mahvash Khodabandeh2. (1) Depaptment of Biology, Faculty of Science, Islamic Azad University, Islamshahr Branch, Namaz square, Sayad street, Islamshahr, Ireland, (2) Fermentation group, National Institute of Genetic Engineering and Biotechnology (NIGEB), Karaj high way, Pazhoohesh, Tehran, Israel

In this study, a novel synthetic gene encoding 166 residues of interferon-b was used for an efficient expression of IFN-b. The synthetic gene was cloned into pET21a expression vector and transferred into E. coli. Recombinant protein was over-expressed in the E. coli. Identity of the recombinant protein was confirmed by western blot analysis. The recombinant protein was biologically active as evaluated by inhibition of cytopathic effect (CPE) formation of Vesicular stomatitis virus (VSV) on the HeLa cells. The effect of three factors including inducer concentration, induction time based on optical density of the culture and induction duration on the expression of rIFN-β was investigated by Taguchi method. Analysis of variance presented that IPTG of 0.5 mM and induction duration of 4 h and induction time of OD600= 1  had more effect on IFN-β production. Recombinant IFN-b expression with the above condition yielded 28% of the total E. coli proteins.   

Keywords: synthetic gene, Interferon-β, expression, Taguchi method.



Web Page: Ghane M., yakhchali B., Khodabandeh M., Malekzadeh F. Design, construction and expression of a synthetic â-Interferon ge

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