High-level expression of a novel liver-targeting fusion interferon with preferred Escherichia coli codon preference and its anti-hepatitis B virus activity in vivo

BMC Biotechnol. 2015 Jun 12;15:54. doi: 10.1186/s12896-015-0177-1.

Abstract

Background: In our previous study, a novel liver-targeting fusion interferon (IFN-CSP) combining IFN α2b with plasmodium region I peptide was successfully constructed. IFN-CSP has significant inhibition effects on HBV-DNA replication in HepG2.2.15 cells. The aim of the present investigation was focused on how to produce high levels of recombinant IFN-CSP and its in vivo anti-hepatitis B virus (HBV) activity.

Methods: A modified DNA fragment encoding IFN-CSP was synthesized according to Escherichia coli (E. coli) preferred codon usage and transformed into E. coli BL21 (DE3) for protein expression. The induction conditions were systematically examined by combining one-factor experiments with an orthogonal test (L(9)(3)(4)). The antigenicity of the purified protein was characterized by western blot analysis. The in vivo tissue distribution were assayed and compared with native IFN α2b. HBV-transgenic mice were used as in vivo model to evaluate the anti-HBV effect of the recombinant IFN-CSP.

Results: The results showed that the E. coli expression system was very efficient to produce target protein.

Conclusion: Our current research demonstrates for the first time that IFN-CSP gene can be expressed at high levels in E. coli through codon and expression conditions optimization. The purified recombinant IFN-CSP showed liver-targeting potentiality and anti-HBV activity in vivo. The present study further supported the application of IFN-CSP in liver-targeting anti-HBV medicines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antiviral Agents / chemistry
  • Antiviral Agents / metabolism*
  • Antiviral Agents / pharmacology
  • Base Sequence
  • Codon / genetics*
  • Escherichia coli / genetics*
  • Hepatitis B virus / drug effects*
  • Interferons / chemistry
  • Interferons / genetics
  • Interferons / metabolism*
  • Interferons / pharmacology
  • Liver / metabolism
  • Liver / virology
  • Mice
  • Mice, Transgenic
  • Molecular Sequence Data
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Recombinant Fusion Proteins / pharmacology

Substances

  • Antiviral Agents
  • Codon
  • Recombinant Fusion Proteins
  • Interferons