鉴定一种新的插入位点，用于PEDV基因组中FMDV VP1的高效表达

Abstract

Porcine epidemic diarrhea virus (PEDV) is an important enteric pathogen that causes severe diarrhea and high mortality in suckling piglets, posing a major threat to the global swine industry. In recent years, viral vectors expressing heterologous protective antigens have emerged as a promising strategy for novel vaccine development. However, whether PEDV can serve as a viral vector for the efficient and stable expression of foreign proteins remains unclear. In this study, a novel insertion site upstream of M in the PEDV genome was identified as a permissive locus for foreign gene insertion using a PEDV reverse genetics platform via yeast-based transformation-associated recombination (TAR) technology. The EGFP reporter system confirmed that the site supported stable foreign gene expression without compromising viral replication. Furthermore, the VP1 of foot-and-mouth disease virus (FMDV), a major immunodominant antigen, was introduced into the site, generating the recombinant virus rPEDV-VP1, which stably maintained the foreign gene not only during serial passaging but also during subsequent in vivo infection in piglets. Collectively, these findings identify a candidate insertion site for efficient expression of foreign antigens in the PEDV genome and provide a technical basis for the development of PEDV-based viral vector platforms.