毕赤酵母表达猪干扰素—γ基因及其抑制蓝耳病毒效果

为了研究和应用猪重组干扰素-γ（rPoIFN-γ)预防和治疗病毒性疫苗，将大白猪PoIFN-γ基因插入到酵母整合载体pHIL-S1，构建了重组GS115工程菌（pHIL-S1/rPoIFN-γ)。经过SDS－PAGE，Western blot分析和抗滤泡性口炎病毒（VSV）活性测定，证实了rPoIFN-γ分子量为18kD，在GS115中的表达量为18％。其抗VSV活性为450－540u/mL。用rPoIFN-γ处理猪肺巨噬细胞系Marc-145后，经细胞病变抑制法（CPE50）测定，rPoIFN-γ可以抑抗蓝耳病病毒（PRRSV）感染。结果显示酵母表达的rPoIFN-γ是有应用价值的抗病毒生物工程制剂。

Expression of porcine interferon-gamma gene in Pichia pastoris and its effect of inhibiting porcine reproductive and respiratory syndrome virus]

In order to develop recombinant porcine Interferon-gamma (rPoIFN-gamma) to prevent porcine viral infection, PoIFN-gamma cDNA lacking the signal peptide was expressed in Pichia pastoris GS115 strain, and the effect of rPoIFN-gamma on porcine reproductive and respiratory syndrome virus (PRRSV) was investigated. The PoIFN-gamma gene was inserted into integrative vector pHIL-S1, and the recombinant GS115 strain (pHIL-S1/PoIFN-gamma) was constructed by homologues recombinant. The rPoIFN-gamma protein was 18 kD with an expressing yield of 18% was assayed by SDS-PAGE and Western blot, respectively. The anti-viral activity of rPoIFN-gamma was in the range of 450-540 u/mL. In addition, the effect of rPoIFN-gamma ant-PRRSV was determined using CPE50 method. The results indicated that high concentration of rPoIFN-gamma could inhibit PRRSV on Marc-145 cell line. The rPoIFN-gamma is a potential drug for prevention and treatment of various kinds of viral pig diseases.