羊口疮病毒口唇和内脏感染株ORFV123基因生物信息学特征与宿主转录响应的比较OA

This study aimed to explore the molecular characteristics of the protein encoded by the ORFV123 gene in the lip and visceral in-fection strains of orf virus(ORFV)and the differences of the protein after being transfected in its host cells.The ORFV123 gene was ampli-fied by PCR technology,and cloned into the pMD-19T vector for sequencing;and its genetic characteristics were analyzed using bioinformat-ic methods.Meanwhile,eukaryotic expression vectors pEGFP-N1-123K and pEGFP-N1-123N were constructed.Subsequently,the eukary-otic expression vectors were transfected into primary goat oral epithelial cells,and their expression levels were measured using Western blot and RT-qPCR,respectively.Finally,the gene expression profiles were analyzed using RNA-seq.The results showed that the full -length of the ORFV123 genes in both the oral and visceral strains was 1 551 bp,encoding 516 amino acids,which belonged to different evolutionary branches and shared with similar physicochemical properties and protein structural characteristics.The three-dimensional structural analysis revealed conformational differences in the C-terminal loop region of the ORFV123 protein between the two strains.The Western blot results showed that the ORFV123 genes of both the strains could be successfully expressed in goat oral epithelial cells.The transcriptome analysis showed that there were 16 differentially expressed genes(DEGs,7 up-regulated and 9 down-regulated)between the orolabial and visceral infection strains of the ORFV,and a total of 1 208 DEGs were identified when the two strains were compared with the blank cell group(806 DEGs in the orolabial strain and 402 DEGs in the visceral strain),which were mainly enriched in signaling pathways such as protein diges-tion and absorption,PI3K-Akt,and AGE-RAGE.These findings provided fundamental insights into the function of the ORFV123 protein and its potential role in viral pathogenesis.