Cloning of vaccine candidates norovirus antigens and investigation of their immunogenities

Objective: This research was carried out in order to obtain recombinant antigenic proteins that can be vaccine candidates of Norovirus and to investigate the immune response that may occur with animal experiments. Material and Methods: The VP1, VP2, p22 and polypeptide (EP123) sequences of the norovirus were amplified in PCR with specific primers. Amplicons were transformed into E. coli BL21 strain after insertion into the pET-30a (+) expression vector. Recombinantly produced proteins were used as antigens for immunization after partial purification with the help of His-tag tail and ammonium sulfate. Antibody response to Norovirus from serum samples of vaccinated mice was determined by indirect ELISA method. Results: The presence of genes transferred by transformation into E. coli was confirmed by sequence analysis. His-tag tail and precipitation with ammonium sulfate were sufficient for partial purification of the proteins. Only the VP1 protein elicited an adequate immune response. Conclusion: In this thesis, in addition to VP1 and VP2 proteins, a polypeptide consisting of epitopes different from previous Norovirus vaccine studies, and a non-structural gene (p22) of Norovirus were produced recombinantly and its immunogenicity was investigated. It was thought that the lack of adequate immunogenic response in recombinant proteins other than VP1 may be related to the dose used, the number of immunizations or the adjuvant administration.