Vector Construction and Immunogenicity of S and N Gene DNA Vaccine for TGEV
[Objective]DNA vaccine vector of S and N genes of porcine transmissible gastroenteritis virus(TGEV)was constructed with the vaccine immunogenicity determined to pave the way for studying,preventing,and controling TGE.[Method]A and D sites on S and N from a TGEV were amplified.The N gene alone as well as the A and D sites fusion were cloned into the vaccine vector pCDNA3.1-His-C.Bioinformatics software was used to predict and analyze the secondary structure,tertiary configuration,subcellular localization,and dominant B cell epitope of S(A-D)and N proteins.The recombinant vectors were transfected into PK-15 cells,and expression distribution of N and the A and D sites fusion detected by indirect immunofluorescence and confocal detection.Mice were immunized with the single or combined recombinant vaccine vector to detect the IgG antibody using indirect ELISA.[Result]The A and D sites of the S were 498 bp and 606 bp,respectively,and the N,1149 bp in length.The nucleic acid vaccine expression vectors p-S(A-D)-His and p-N-His for the A and D sites(fusion)and N were constructed.Bioinformatics software predicted that,when TGEV infected the host cells,N protein was mainly located in the nucleus and mitochondria and S(A-D)largely in the cytoplasm and mitochondria,while S(A-D)had 7 and N,8 dominant B cell epitopes.All p-S(A-D)-His and p-N-His were successfully expressed in PK-15 cells distributed in the nucleus and cytoplasm.The immunized mice showed an effect of immunity in the order of p-N-His＞p-S(A-D)-His+p-N-His＞p-S(A-D)-His.[Conclusion]The DNA vaccine vectors of S and N of TGEV were successfully constructed.Strong specific antibodies were generated in lab mice after the immunization.
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