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Secretory expression of porcine interferon-gamma in baculovirus using HBM signal peptide and its inhibition activity on the replication of porcine reproductive and respiratory syndrome virus

文献类型： 外文期刊

作者： Wang, Yan-Bin 1 ; Wang, Zhen-Ya 1 ; Chen, Hong-Ying 1 ; Cui, Bao-An 1 ; Wang, Ya-Bin 1 ; Zhang, Hong-Ying 1 ; Wang, Ru 1 ;

作者机构： 1.Henan Agr Univ, Key Lab Anim Derived Food Safety Henan Prov, Coll Husb & Vet, Zhengzhou 450002, Henan, Peoples R China

2.Henan Acad Agr Sci, Zhengzhou 450002, Henan, Peoples R China

关键词： Porcine;Interferon-gamma;Baculovirus;HBM signal peptide;Inhibition activity;PRRSV

期刊名称：VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY （ 影响因子：2.046； 五年影响因子：2.217 ）

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收录情况： SCI

摘要： The gene sequence encoding mature porcine interferon-gamma (PoIFN- gamma ) fused with a C-terminal 6x histidine tag was cloned into the baculovirus pFastBac(TM) Dual vector of the Bac-to-Bac Baculovirus expression system under the control of PH promoter. The authentic signal sequence of porcine interferon-gamma was substituted with the honeybee melittin (HBM) signal sequence, and expressed in insect cells. The recombinant proteins were detected by SDS-PAGE and immunofluorescence assay. The nickel affinity column purified recombinant porcine interferon-gamma with HBM signal peptide (rPoIFN- gamma H) was shown to be a 19kDa protein as confirmed by Western blot analysis. The recombinant PoIFN- gamma H was shown to have cytokine activity, inhibiting the cytopathic effect of vesicular stomatitis virus (VSV) in PK-15 cells at about 1.07x10 super(6)U/mL. The 2 super(-) super(7) dilution of the rPoIFN- gamma H in culture supernatant protected the MARC-145 cells from the cytopathic effect caused by 100TCID sub(5) sub(0) of porcine reproductive and respiratory syndrome virus.

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