Cloning and expression recombinant human interferon-β-1b

Abstract

Recombinant human Interferon-  - 1 b (IFN-  - 1 b) is a non-glycosylated IFN-  that was found to be effective against multiple sclerosis. More recently, expression of IFN-!-1b in Escherichia coli (E. coli) has been reported; however, problems due to low yields and poor stability were described. This study was aim to clone and express of recombinant IFN-!-1b in E. coli using a novel synthetic gene with preferred codon usage of E. coli under the control of T7 promoter. Protein expression was induced with Isopropyl-thiogalactoside (IPTG) and lactose. The level of expression was analyzed by SDS-PAGE, Western blot, Bradford and ELISA. The effect of two factors including inducer concentration, and length of the induction on the expression of IFN-!-1b was investigated. As a result, we have successfully cloned the E. coli BL21 (DE3) stain that harbors gene encoding for rhIL-29. Results from SDS-PAGE, Western blot and ELISA show that IPTG of 1 mM and induction duration of 6h had more effect on IFN-!-1b production. IFN-  - 1b expression with the above condition yielded 17.39% of the total cell protein. Besides, under the induction of 0.25% lactose and 6 hours of incubation, IFN-  - 1 b production also achieved a high efficiency 13.4% of total cell protein, which promises for an effective way for industrial manufacture. Keywords: Expression, Interferon-β-1b, E. coli BL21 (DE3), T7 promoter, induction.