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中国巴马小型猪干扰素-αω的基因克隆、原核表达及抗病毒活性。

Gene cloning, prokaryotic expression and antiviral activities of interferon-αω from Chinese Bama miniature pigs.

机构信息

The College of Veterinary Medicine, Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Diseases and Zoonoses, Yangzhou University, Yangzhou, 225009, China.

The College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, China.

出版信息

Vet Res Commun. 2022 Feb;46(1):59-66. doi: 10.1007/s11259-021-09829-9. Epub 2021 Sep 28.

Abstract

Porcine interferon (PoIFN) complex represents an ideal model for studying IFN evolution that resulted from viral pressure during domestication. IFN-αω is an emergent subtype of type I IFNs which has been primarily characterized in domestic pigs. In this study, the PoIFN-αω cDNA was cloned from Chinese Bama miniature pigs by RT-PCR, and its tissue expression profile was analyzed by real-time RT-PCR. The cDNA was expressed in Escherichia coli as a His-tagged protein and purified by nickel affinity chromatography. The antiviral activities of recombinant PoIFN-αω (rPoIFN-αω) against four different pig viruses were measured using cytopathic effect (CPE) inhibition assay. Although the PoIFN-αω sequence of Bama miniature pigs was identical to that of domestic pigs, the tissue expression profiles differed significantly between the two pig species. The rPoIFN-αω showed dose-dependent pre-infection antiviral activities against porcine pseudorabies virus, vesicular stomatitis virus and porcine reproductive and respiratory syndrome virus, but not against porcine circovirus type 2. When used as treatment post infection with the three viruses, rPoIFN-αω showed the efficacy in decreasing CPE in the infected cells in a time-dependent manner. Therefore, the expressed rPoIFN-αω could be used as an antiviral agent against pig virus infections.

摘要

猪干扰素 (PoIFN) 复合物是研究干扰素进化的理想模型,这种进化是由驯化过程中的病毒压力导致的。IFN-αω 是 I 型干扰素的新兴亚型,主要在驯化的家猪中得到了表征。在这项研究中,通过 RT-PCR 从中国巴马小型猪中克隆了 PoIFN-αω cDNA,并通过实时 RT-PCR 分析了其组织表达谱。通过将 cDNA 在大肠杆菌中表达为 His 标记的蛋白,并通过镍亲和层析进行纯化。使用细胞病变效应 (CPE) 抑制测定法测量了重组猪干扰素-αω (rPoIFN-αω) 对四种不同猪病毒的抗病毒活性。尽管巴马小型猪的 PoIFN-αω 序列与家猪相同,但这两个猪种的组织表达谱存在显著差异。rPoIFN-αω 对猪伪狂犬病病毒、水疱性口炎病毒和猪繁殖与呼吸综合征病毒具有剂量依赖性的感染前抗病毒活性,但对猪圆环病毒 2 型没有活性。当用作三种病毒感染后的治疗时,rPoIFN-αω 表现出以时间依赖性方式降低感染细胞中 CPE 的功效。因此,表达的 rPoIFN-αω 可用作抗猪病毒感染的抗病毒剂。

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