Expression,Purification And Immunogenicity Identification Of Recombinant Antigens Of Rotavirus And Norovirus

Rotavirus is the main pathogen causing diarrhea in infants and young children.Norovirus can cause diarrhea in infants and young children.It can also cause adult infection and cause gastroenteritis.The two viruses mainly infect intestinal epithelial cells,causing cell damage and causing diarrhea.Clinical symptoms such as vomiting cause a large number of direct or indirect economic losses and even lead to a large number of child deaths worldwide.Vaccination is the main means of preventing and controlling rotavirus and Norovirus infection.Rotavirus vaccine has been used in clinical practice for many years,which can significantly reduce the symptoms of rotavirus infection and the symptoms caused;it is difficult to achieve the isolation and culture of Norovirus in vitro.No Norovirus vaccine has been marketed,and the Norovirus vaccine under research is mainly genetically engineered vaccine.Therefore,we base ourselves on genetic engineering vaccines,and the main antigenic epitopes of the two viruses are merging together to construct an expression vector for expression.Purification and immunogenicity studies are intended to be a preliminary work for future possible joint vaccine development.Objectives:The purpose of this paper is to express the core epitope of the VP7 gene of G1(ZTR-68)and G9(ZTR-18)rotavirus and the core epitope of G?.4 Norovirus VP1 gene.It is displayed on the surface of hepatitis B core antigen particles.To establish a theoretical basis for the establishment of a combined subunit vaccine carrying rotavirus and norovirus core antigen based on hepatitis B core antigen.Methods:A plasmid for the tandem recombinant antigen of G1 type and G9 type rotavirus carrying the HBV core epitope,a recombinant antigen of the G?.4 Norovirus VP1 gene,G1 type and G9 type and G?.4 type A plasmid for the recombinant antigen of the Norovirus VP1 gene,which is linked by a linker peptide GGGGS in the middle of different amino acid segments,and then induced by IPTG to express the recombinant protein in E.coli and purify the epitope containing the above antigen.Recombinant proteins were identified for various immune-related experiments and immunized to detect their immunogenicity.Results:Successfully constructed a tandem recombinant antigen plasmid of G1 type and G9 type rotavirus,a recombinant antigen plasmid of G?.4 Norovirus VP1 gene,G1 type rotavirus and G9 type rotavirus and G?.4 type Nuru The plasmid of the tandem recombinant antigen of the virus was transformed into E.coli DH5a competent bacteria,and the recombinant protein containing the above epitope was efficiently expressed by IPTG.The experiment showed that the protein was recognized by the specific virus antibody and was immunized with the recombinant protein antigen.Subsequent mouse serotyping indicates that the recombinant protein can elicit a high level of specific antibody responseConclusions:Tandem recombinant antigen plasmid of G1 type and G9 type rotavirus,plasmid of recombinant antigen of G?.4 Norovirus VP1 gene,tandem of G1 type rotavirus and G9 type rotavirus and G?.4 Norovirus Plasmids that recombine antigens induce high levels of persistence of specific antibody responses.