Over expression of a synthetic gene encoding interferon lambda using relative synonymous codon usage bias in Escherichia coli

Pak J Pharm Sci. 2013 Nov;26(6):1181-8.

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.

MeSH terms

  • Cloning, Molecular
  • Codon*
  • Escherichia coli / genetics*
  • Gene Expression
  • Genes, Synthetic*
  • Interferons / genetics*

Substances

  • Codon
  • Interferons