title: 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.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: School of Basic Courses, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, People's Republic of China. luxuemei605@163.com.|Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, People's Republic of China. luxuemei605@163.com.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Lu
authors.affiliation: School of Basic Courses, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, People's Republic of China.|Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, People's Republic of China.
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Basic Courses, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, People's Republic of China.|Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, People's Republic of China.
authors.forename: Xiaobao
authors.initials: X
authors.lastname: Jin
authors.affiliation: School of Basic Courses, Guangdong Pharmaceutical University, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, People's Republic of China. zhujiayong1020@163.com.|Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, 280 Wai Huan Dong Road, Guangzhou Higher Education Mega Center, Guangzhou, People's Republic of China. zhujiayong1020@163.com.
authors.forename: Jiayong
authors.initials: J
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D001483:Base Sequence; D003062:Codon / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D006515:Hepatitis B virus / Q000187:drug effects*; D007372:Interferons / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D008099:Liver / Q000378:metabolism / Q000821:virology; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D000998:Antiviral Agents; D003062:Codon; D011993:Recombinant Fusion Proteins; D007372:Interferons