猪干扰素α、β和γ的融合表达及活性

doi:10.3864/j.issn.0578-1752.2012.09.020

Abstract

Abstract: 【Objective】The research aimed to exploit high efficient porcine gene engineering antiviral agents to prevent and control the porcine viral diseases and tumor diseases.【Method】Three fusion genes of PoIFN-α/β, PoIFN-α/γ and PoIFN-β/γ were constructed by linking any two mature peptide genes of the PoIFN-α, PoIFN-β and PoIFN-γ by SOE-PCR method. Then, these fusion genes were subcloned into prokaryotic expression vector pET-30a for expression in E. coli BL21(DE3). Subsequently, the bioactivities of the fusion proteins rPoIFN-α/β, rPoIFN-α/γ and rPoIFN-β/γ were comparatively analyzed with non-fusion proteins rPoIFN-α, rPoIFN-β and rPoIFN-γ by the cytopathic effect (CPE) inhibition assay and the porcine lymphocyte transformation assay.【Result】The fusion genes of PoIFN-α/β, PoIFN-α/γ and PoIFN-β/γ were successfully constructed and expressed in E. coli. After purified and refolded from inclusion bodies, three fusion proteins of rPoIFN-α/β, rPoIFN-α/γ and rPoIFN-β/γ with well biological activity were obtained successfully. The results of the cytopathic effect (CPE) inhibition assay indicated that all these three kinds of fusion proteins, which demonstrated more significant antiviral activities for PRRSV or VSV than non-fusion proteins rPoIFN-α, rPoIFN-β and rPoIFN-γ not only on Marc-145 but also on PK-15, exhibited increased antiviral potencies. The effects of rPoIFN-α/β, rPoIFN-α/γ and rPoIFN-β/γ on the porcine lymphocyte transformation were detected by the MTT method. The results suggested that both rPoIFN-α/γ and rPoIFN-β/γ could effectively stimulate the transformation of porcine lymphocyte, had obvious immunoregulatory activities, but rPoIFN-α/β had no obvious effect. So, there are synergistic immunoregulatory effects between typeⅠand type Ⅱ PoIFNs, and none between PoIFNs of type Ⅰ.【Conclusion】 All these three fusion PoINFs in study, which exhibited the prominent abilities of antivirus, could be used to prevent and control the porcine viral diseases and tumor diseases. And the rPoIFN-α/γ and rPoIFN-β/γ also had stronger immune enhancement and could be expected to exploit into new-type immunopotentiators.

Key words: porcine interferon, fusion expression, antiviral activity, cytopathic effect inhibition, porcine lymphocyte transformation

ZHENG Ming, BIAN Chuan-Zhou, WANG Lao-Qi, WANG Yong-Fen. Study on the Fusion Expression and Bioactivity of Porcine Interferon α, β and γ[J].Scientia Agricultura Sinica, 2012, 45(9): 1840-1847.

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