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Updated: May 22, 2026
Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing
Published on: January 7, 2019
[Construction of a recombinant eukaryotic expression vector containing HSV 2gD and its immune responses]
Feng-ping Jiao1, Li Liu, Yu Wang
1Department of Aetiology, Taishan Medical University, Taian, China. fpjiao@tsmc.edu.cn
Aim:
To construct and express pcDNA3.1-IL-18-HSV P6 recombinant DNA vaccine, and to observe the immune responses of pcDNA3.1-IL-18-HSV P6 genetic vaccine.
Methods:
Genes of P6 and IL-18 were cloned into the eukaryotic expression vector pcDNA3.1(-) respectively, and confirmed by enzyme digestion and sequence analysis, the recombinant plasmid pcDNA3.1-IL-18-HSVP6 was transformed into CHO cells. The expressed protein was characterized by indirect immunofluorescence and Western blotting. The recombinant plasmid pcDNA3.1-IL-18-HSVP6 was used to inoculate BALB/c mice by intramuscular injection for three times, once a week. One week after the last vaccination, the levels of specific IgG antibody, IFN-γ and IL-18 were detected by ELISA; One month after the last vaccination, spleen cells of vaccinated mice were separated and proliferation of spleen lymphocytes were detected by MTT assay.
Results:
Recombinant pcDNA3.1-IL-18-HSV P6 plasmid was confirmed by enzyme digestion and DNA sequencing. After inoculated by pcDNA3.1-IL-18-HSV P6 vaccine, the mice could produce higher level of splenocytes proliferation and secrected higher level of IFN-γ, IL-18 and specific antibody.
Conclusion:
DNA vaccine of pcDNA3.1-IL-18-HSV P6 has been successfully constructed, and it can effectively induce humoral and cellular immune responses, which provided a basis for constructing new type of DNA vaccine.
