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Indexed/Abstracted in: EMBASE, Science Citation Index Expanded (SciSearch), Scopus
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Mohammad A. MOBASHER 1, 2, Younes GHASEMI 1, 2, Nima MONTAZERI-NAJAFABADY 1, 2, Abbas TAHZIBI 3
1 Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Fars, Iran; 2 Department of Pharmaceutical Sciences Research Center, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Fars, Iran; 3 Food & Drug organization Ministry of Health, Tehran, Iran
BACKGROUND: Interferon beta 1-b (IFN β 1-b) has antiviral, anti-inflammatory, antiproliferative and immunomodulatory properties. It is a nonglycosylated protein produced by the cells of Escherichia coli (E. coli) and has many applications in treatments of hepatitis, glyoma, genital condylomata acuminate, multiple sclerosis (MS), melanoma and rheumatoid arthritis. Many studies have been done to overexpress this valuable protein. In this work the patterns of IFN β 1-b expression, in active (soluble) and inactive (inclusion body) forms were evaluated.
METHODS: Two types of expression vectors were chosen. pET25b, that includes pelB signal peptide which makes protein as secretable forms that migrate to periplasmic space, and pET15b with a His-tag sequence at N-terminal of the IFN β 1-b gene which tends to express recombinant proteins in the form of insoluble aggregates called inclusion bodies. IFN β 1-b was transformed into E. coli BL21 (DE3), Rosetta and JM109 and after induction with Isopropyl β-D-1-thiogalactopyranoside (IPTG), the amounts of protein expression were read through density of IFN β 1-b bonds by ImageJ software as a percent of total cell protein.
RESULTS: This study showed that expression of IFN β 1-b in soluble form was too low to be detected by SDS-PAGE while production in form of inclusion bodies (IBs) resulted in an expression about 30% of total cell proteins. The most expressions were belonging to E. coli BL21 (DE3) and Rosetta and the least to JM109.
CONCLUSION: In the case of toxic proteins to E. coli cells, like IFN β 1-b, expression of soluble protein with correct folding which is active, can affect host cell and reduce the yield of production. But inclusion bodies are a way to overcome this problem. Besides, expression of IFN β 1-b in this form is simple and cost benefit.