Porcine teschovirus (PTV) is a causative agent of polio, encephalomyelitis, reproductive disorders, and gastrointestinal and respiratory diseasesin swine. In this study, the PTV-2 GX/2020 strain was isolated from pig intestinal tissue using ST cells. Phylogenetic analysis of VP1 nucleotide sequences indicated that the GX/2020 strain was closely related to PTV-2 (86%). Furthermore, a full-length cDNA infectious clone of GX/2020 was constructed using seamless ligation technology. The rescued virus was had a genome sequence largely consistent with the parental virus sequence and had similar growth characteristics. Overall, the PTV-2 virus successfully isolated in this study and the employed reverse genetic platform have laid a foundation for studying the pathogenic mechanism of porcine Teschovirus.