PDCoV E Protein prokaryotic expression and phylogenetic analysis
The eukaryotic expression of E protein in Porcine Delta Coronavirus strain was analyzed and phylogenetic analysis was performed,which laid a foundation for the further exploration of the biological characteristics of E gene in PDCoV and the establishment of diagnostic methods.In this study,PDCoV E gene was amplified by reverse transcription using PDCoV-HNZMD RNA as template,ligated and transformed to construct the eukaryotic expression recombinant plasmid pcDNA3.1-E.After transfected with HEK-293T cells,the gene expression was detected by IFA and western blot.The target gene of PDCoV E protein was successfully obtained,and the eukaryotic vector pcDNA3.1-E was successfully constructed.The results of IFA and Western blot showed that the recombinant eukaryotic expression plasmid could correctly express,the proportion of amino acid homology was 92.9%-100%.The genetic evolution tree analysis showed that PDCOV-HNZMD was most closely related to the Guangxi strain(OQ736717).In this study,the eukaryotic expression plasmid of PDCoV-E protein was successfully constructed and verified by IFA and WB,which provides a reference for the subsequent research on the pathogenic mechanism.
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