title: Cloning and Expression of Chinese Duck Interferon-gamma Gene.
abstract: The efficacy of cytokine therapy has been demonstrated in several viral diseases. Interferon-gamma is a cytokine that has potent antiviral property and immunomodulatory activity. To investigate the role of IFN-gamma in viral clearance during natural infection and to define the antiviral mechanism, DHBV-infected ducks was used as an animal model. To clone, express, and develop the method of quantifying DuIFN-gamma gene transcription and expression, DuIFN -gamma cDNA was amplified by RT-PCR from PHA stimulated duck PBMC. Recombinant plasmid expressing DuIFN-gamma was used to transfect COS-7, and the cell culture supernatant was analyzed by CPE inhibitory assay and MTT methods to determine the antiviral titer of IFN-gamma. The GST-DuIFN-gamma fusion protein was expressed in E.coli and purified using the GST sepharose 4B. Results indicated that the supernatant collected from COS-7 cells transfected with DuIFN-gamma cDNA was able to prevent duck fibroblasts from VSV induced CPE in a dose dependent manner. An anti-DuIFN-gamma antibody neutralized this antiviral activity.
authors.affiliation: Department of Molecular Virology, Medical Center of Fudan University, Shanghai 200032, China. d-qu@usa.net
authors.forename: J E
authors.initials: JE
authors.lastname: Long
authors.forename: L N
authors.initials: LN
authors.lastname: Huang
authors.forename: W Y
authors.initials: WY
authors.lastname: Wang
authors.forename: M J
authors.initials: MJ
authors.lastname: Cheng
authors.forename: Y M
authors.initials: YM
authors.lastname: Wen
authors.forename: Z H
authors.initials: ZH
authors.lastname: Yuan
authors.forename: D
authors.initials: D
authors.lastname: Qu