Evaluation of the transcriptional regulatory efficacy of transcription regulatory sequences of porcine epidemic diarrhea virus

•This is the first report to show that exogenous gene could be inserted between N gene and 3’ UTR of PEDV.•Insertion of exogenous gene between N and 3’ UTR of PEDV does not influence the PEDV replication characteristics.•The TRS of M protein gene produced the highest level of EGFP in all TRSs of PEDV. Porcine epidemic diarrhea virus (PEDV) is a highly contagious enteropathogenic coronavirus causing severe watery diarrhea and high mortality in piglets. In order to investigate the role of the transcription regulatory sequences (TRSs) in regulation of gene expression and replication of PEDV, the enhanced green fluorescent protein (EGFP) gene, under control of different TRSs of PEDV, were inserted between the N gene and 3′ UTR of the PEDV genome using a reverse genetic system. The EGFP expression from different chimeric PEDVs was analyzed for each TRS. TRSs of all the structural and accessory protein genes of PEDV positively regulate EGFP expression at different levels, and the TRS of M protein gene produced the highest level of EGFP. Moreover, this is the first study to show that exogenous gene could be inserted between N gene and 3’ UTR of PEDV, and the EGFP insertion had no effect on PEDV replication. Taken together, our study enriched the information of PEDV TRSs on gene expression and replication of PEDV.