title: [Cloning, prokaryotic expression of chicken interferon-alpha gene and study on antiviral effect of recombinant chicken interferon-alpha].
abstract: The full length of chicken interferon alpha (ChIFN-alpha) gene was amplified by the polymerase chain reaction (PCR) from total liver genome of Sanhuang meat-chicken and sequenced. The amplified gene was about 582bp. The coding region for mature protein (489bp) was subcloned into pET-28a(+). The recombinant plasmid pET-28a(+)-IFNalpha was identified by enzyme digestion and DNA sequencing. Data of SDS-PAGE and Western-blot indicated that a 22kD fusion protein was expressed in the form of inclusion bodies with good immunity. The purity of inclusion bodies was above 70% and that of protein purified by nickel affinity chromatography was 95%. The recombinant protein could inhibit H9N2 avian influenza virus (H9N2 AIV) replication on chick embryo fibroblast. 2 microg of recombinant IFN-alpha could completely protect Chick embryo from H9N2 AIV infection. The recombinant IFN-alpha can also delay Newcastle disease virus (NDV) replication on chick embryo for 12 approximately 48h. Chicken administered recombinant IFN-alpha can resist the H9N2 AIV infection. The bioactivities of recombinant IFN-alpha purified by affinity chromatograph were 20 times higher than that of inclusion bodies.
authors.affiliation: Lab of Animal Virus, State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Wei
authors.forename: Gui-Qing
authors.initials: GQ
authors.lastname: Peng
authors.forename: Mei-Lin
authors.initials: ML
authors.lastname: Jin
authors.forename: Yu-Dong
authors.initials: YD
authors.lastname: Zhu
authors.forename: Hong-Bo
authors.initials: HB
authors.lastname: Zhou
authors.forename: Hong-Yan
authors.initials: HY
authors.lastname: Guo
authors.forename: Huan-Chun
authors.initials: HC
authors.lastname: Chen
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D015153:Blotting, Western; D002642:Chick Embryo; D003001:Cloning, Molecular; D053127:Influenza A Virus, H9N2 Subtype / Q000187:drug effects; D007370:Interferon Type I / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology*; D016898:Interferon-alpha / Q000235:genetics*; D009522:Newcastle disease virus / Q000187:drug effects; D010957:Plasmids; D011994:Recombinant Proteins
chemical_list: D000998:Antiviral Agents; D007370:Interferon Type I; D016898:Interferon-alpha; D011994:Recombinant Proteins