Construction and expression of a recombinant plasmid containing the porcine epidemic diarrhea virus S1 gene delivered by Lactobacillus

摘要

[Objective] To construct a recombinant lactobacillus capable of expressing the porcine epidemic diarrhea virus S1 gene, and to lay a foundation for subsequent research on oral vaccination against porcine epidemic diarrhea virus. [Methods] The bioinformatics software Protean was used to analyse the antigenicity of the PEDV S1 protein and the 307 amino acid neutralising epitope was screened and sent to Sangon Biotech Co., Ltd for codon optimisation. The plasmid was ligated to the pUC-57 clone plasmid to form the pUC57-Sl recombinant plasmid. The pVE5523 vector and the pUC57-S1 recombinant plasmid were enzymatically digested and the target gene was spliced into the final vector pVE5523 to obtain a recombinant plasmid pVE5523-Sl, which was electroporated and transformed into Lactobacillusplantarum to obtain a recombinant Lactobacillus plantarum. The recombinant Lactobacillus plantarum was identified by western blot to demonstrate whether the target gene was expressed. [Result] The recombinant plasmids pUC57-Sl and pVE5523-Sl were successfully constructed using Lactobacillus plantarum as a template and the recombinant plasmids pUC57-Sl and pVE5523-Sl were successfully transformed. Western blot analysis showed that the recombinant Lactobacillus successfully expressed a recombinant protein with a relative molecular mass of approximately 34 kD.