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猪γ-干扰素基因在毕赤酵母中的表达及其对猪繁殖与呼吸综合征病毒的抑制作用

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

作者信息

Wan Jian-Qing, Wu Wen-Xue, Xia Chun

机构信息

College of Veterinary Medicine, Chinese Agricultural University, Beijing 100094, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2002 Nov;18(6):683-6.

Abstract

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.

摘要

为研发重组猪干扰素-γ(rPoIFN-γ)以预防猪病毒感染,在毕赤酵母GS115菌株中表达缺失信号肽的PoIFN-γ cDNA,并研究rPoIFN-γ对猪繁殖与呼吸综合征病毒(PRRSV)的作用。将PoIFN-γ基因插入整合载体pHIL-S1,通过同源重组构建重组GS115菌株(pHIL-S1/PoIFN-γ)。分别通过SDS-PAGE和Western blot检测,rPoIFN-γ蛋白为18 kD,表达量为18%。rPoIFN-γ的抗病毒活性在450 - 540 u/mL范围内。此外,采用CPE50法测定rPoIFN-γ抗PRRSV的效果。结果表明,高浓度的rPoIFN-γ可在Marc-145细胞系上抑制PRRSV。rPoIFN-γ是预防和治疗多种猪病毒性疾病的潜在药物。

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