Atypical porcine pestivirus (APPV) is a novel pestivirus causing congenital tremor (CT) type AII in piglets and exhibiting a broad geographical distribution. Lack of an operating system for the viral genome is one of bottlenecks which restrict further research on pathogenesis and gene functions of APPV. Reverse genetics system (RGS) is a feasible solution to this bottleneck problem, but, to-date, no RGSs have been developed for APPV. Here, for the first time, recombinant APPV CH-GD2017 were rescued using in vitro and intracellular transcription systems and the virons were observed via transmission electron microscopy. As the process of in vitro transcription is time-consuming and inefficient, a full-length cDNA clone in an intracellular transcription was further constructed using an RNA polymerase II system. Then, the rescued virus was identified via RT-PCR detection, indirect immunofluorescent assay, and transmission electron microscopy. Development of the RGS for APPV will provide an important tool for further research on this newly emerging pathogen.

中文翻译：

---

使用体外和细胞内转录系统开发新兴非典型猪瘟病毒的反向遗传学系统。

非典型猪瘟病毒 (APPV) 是一种新型瘟病毒，可引起仔猪先天性震颤 (CT) 型 AII，具有广泛的地理分布。缺乏病毒基因组操作系统是制约APPV发病机制和基因功能进一步研究的瓶颈之一。反向遗传系统 (RGS) 是解决这一瓶颈问题的可行方法，但迄今为止，尚未为 APPV 开发 RGS。在这里，首次使用体外和细胞内转录系统拯救了重组 APPV CH-GD2017，并通过透射电子显微镜观察了病毒体。由于体外转录过程耗时且效率低下，因此使用RNA聚合酶II系统进一步构建了细胞内转录的全长cDNA克隆。然后，通过RT-PCR检测、间接免疫荧光分析和透射电镜鉴定获救的病毒。为 APPV 开发 RGS 将为进一步研究这种新出现的病原体提供重要工具。