Author/Authors :
Akhtar, Hashaam National University of Sciences and Technology - Atta-Ur-Rahman School of Applied Biosciences, Pakistan , Akhtar, Samar Riphah International University - Riphah Institute of Pharmaceutical Sciences, Pakistan , Jan, Umer University of Balochistan - Department of Pharmacy, Pakistan , Khan, Azka National University of Sciences and Technology - Atta-Ur-Rahman School of Applied Biosciences, Pakistan , Zaidi, Najam us Sahar Sadaf National University of Sciences and Technology - Atta-Ur-Rahman School of Applied Biosciences, Pakistan , Qadri, Ishtiaq saudi arabia - Head Medical Biotechnology, King Fahd Medical Research Center, Saudi Arabia
Abstract :
Interferon Lambda (IFN-λ) is a type III interferon which belongs to a novel family of cytokines and possesses antiviral and antitumor properties. It is unique in its own class of cytokines; because of the specificity towards its heterodimer receptors and its structural similarities with cytokines of other classes. This renders IFN-λ a better choice for the treatment against many diseases including viral hepatitis and human coronavirus (HCoV-EMC). The present study describes a computational approach known as relative synonymous codon usage (RSCU); used to enhance the expression of IFN-λ protein in a eukaryotic expression system. Manually designed and commercially synthesized IFN-λ gene was cloned into pET-22b expression plasmid under the control of inducible T7-lac promoter. Maximum levels of IFN-λ expression was observed with 0.4 mM IPTG in transformed E. coli incubated for 4 hours in LB medium. Higher concentrations of IPTG had no or negative effect on the expression of IFN-λ. This synthetically over expressed IFN-λ can be tested as a targeted treatment option for viral hepatitis after purification.
