Cloning, prokaryotic expression of chicken interferon-alpha gene and study on antiviral effect of recombinant chicken interferon-alpha / 生物工程学报
Chinese Journal of Biotechnology
; (12): 737-743, 2006.
Article
ي Zh
| WPRIM
| ID: wpr-286217
المكتبة المسؤولة:
WPRO
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.
النص الكامل:
1
الفهرس:
WPRIM
الموضوع الرئيسي:
Antiviral Agents
/
Pharmacology
/
Plasmids
/
Recombinant Proteins
/
Newcastle disease virus
/
Interferon Type I
/
Blotting, Western
/
Interferon-alpha
/
Cloning, Molecular
/
Influenza A Virus, H9N2 Subtype
المحددات:
Animals
اللغة:
Zh
مجلة:
Chinese Journal of Biotechnology
السنة:
2006
نوع:
Article