Expression and Antiviral Activity Detection of Porcine Interferon -gamma in Silkworm

doi:10.13560/j.cnki.biotech.bull.1985.2016.01.024

Abstract

Abstract: Interferon is used for threating viral infectious diseases and enhancing vaccine efficacy in stockbreeding. In this study, recombinant porcine interferon-gamma(PoIFNγ)was expressed in silkworm by using Baculovirus expression vector system. Based on the published sequences, the gene of PoIFNγ was artificially synthesized after codon optimization. The optimized gene was firstly cloned into pVL1393 transfer vector of Baculovirus, with virus BmBacmid DNA which was co-transfected into BmN cell line, and recombinant BmNPV was obtained. The PoIFNγ gene was in the downstream of polyhedrosis gene promoter in BmNPV. The silkworm larva infected with recombinant BmNPV gained the expression of PoIFNγ that was detected by Western blotting. The activity of interferon was measured by using micro-cytopathic method that uses interferon to inhibit the VSV-GFP infecting VERO cells. The results showed that the titer of interferon was over 6×10⁵ IU/mL. The activated PoIFNγ was successfully expressed in silkworm larvae.

Key words: porcine interferon-gamma, silkworm, Baculovirus expression system, antiviral activity

LIU Xing-jian, LI Hao-yang, HU Xiao-yuan, ZHANG Zhi-fang, YI Yong-zhu, LI Yi-nv. Expression and Antiviral Activity Detection of Porcine Interferon -gamma in Silkworm[J]. Biotechnology Bulletin, 2016, 32(1): 144-148.

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