Prokaryotic Expression of Recombinant Interferon IFNγ of Grass Carp (Ctenopharyngodon idella) and Its Anti-GCRV Effects
YUAN Xue-Mei1, XU Ling-Wei2, HUANG Lei1, HAO Gui-Jie1, LIN Ling-Yun1, PAN Xiao-Yi1, YAO Jia-Yun1, SHEN Jin-Yu1*
1 Key Laboratory of Healthy Freshwater Aquaculture, Ministry of Agriculture and Rural Affairs/Key Laboratory of Fish Health and Nutrition of Zhejiang Province/Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China;
2 Hangzhou Xiaoshan Aquatic Station of Agricultural and Forestry Technology Extension Center, Hangzhou 311225, China
Abstract：In recent years, grass carp hemorrhagic disease caused by Grass carp reovirus (GCRV) has caused serious losses to the grass carp industry. In order to explore the immune protective effects of recombinant interferon on grass carp and its mechanism, the prokaryotic expression vector PET-30a-rCiIFNγ of Ctenopharyngodon idella interferon γ (IFNγ) was constructed and transferred into Transetta (DE3). The recombinant protein rCiIFNγ inducted by IPTG and purified by nickel column chromatography. The antiviral activity of rCiIFNγ in vitro was detected by cytopathic effect (CPE) observation. The protective efficacy of rCiIFNγ in vivo was evaluated by challenge experiment. The results showed that the molecular weight of rCiIFNγ is about 23kD, and the antiviral titer of purified rCiIFNγ was 2.8×103 U/μg. rCiIFNγ could not only inhibit cytopathic effect induced by GCRV in vitro, but also induced the expression of IFN-stimulated genes (IFN, IRF1, IRF7, Mx, PKR, STAT3 and IFNr) in liver, spleen, kidney and intestine, and significantly reduced the mortality of C. idellus challenged with GCRV. These results indicated that the recombinant interferon IFNγ fusion protein rCiIFNγ was effective in protecting grass carp against GCRV infection. This study provides a theoretical basis for the antiviral application of grass carp interferon γ.
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