Construction and Immunogenicity of S and N gene DNA Vaccine Vector for TGEV

CHENG Weiwei; RONG Weizhong; YANG Ming; LI Yuanxin; ZHAO Zihui; CHEN Boxiang; WANG Jia; ZHOU Yao

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CHENG W W, RONG W Z, YANG M, et al. Construction and Immunogenicity of S and N gene DNA Vaccine Vector for TGEV [J]. Fujian Journal of Agricultural Sciences，2024，39（9）：1−9

Citation:

CHENG W W, RONG W Z, YANG M, et al. Construction and Immunogenicity of S and N gene DNA Vaccine Vector for TGEV [J]. Fujian Journal of Agricultural Sciences，2024，39（9）：1−9

CHENG W W, RONG W Z, YANG M, et al. Construction and Immunogenicity of S and N gene DNA Vaccine Vector for TGEV [J]. Fujian Journal of Agricultural Sciences，2024，39（9）：1−9

Citation:

CHENG W W, RONG W Z, YANG M, et al. Construction and Immunogenicity of S and N gene DNA Vaccine Vector for TGEV [J]. Fujian Journal of Agricultural Sciences，2024，39（9）：1−9

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Construction and Immunogenicity of S and N gene DNA Vaccine Vector for TGEV

Gansu Institute of Animal and Veterinary Science , Pingliang, Gansu　744000, China

Received Date: 2024-06-03
Rev Recd Date: 2024-08-23

Available Online: 2024-10-29

Abstract

Abstract

Objective In order to provide a technical support and basic data for the prevention and control of porcine transmissible gastroenteritis and the DNA vaccine research, the DNA vaccine vector of S and N genes of porcine transmissible gastroenteritis virus was constructed and its immunogenicity was tested. Method The A site and D site from the S gene and N gene was amplified from transmissible gastroenteritis virus and the N gene (alone) and 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 composition, tertiary configuration, subcellular localization and dominant B cell epitope of S(A-D) protein and N protein. The recombinant vectors were transfected into PK-15 cells, and expression distribution of the N gene (alone) and the A and D sites (fusion) was detected by indirect immunofluorescence and confocal detection. The recombinant vaccine vector was used to immunize mice alone or in combination, and the IgG antibody level was detected by indirect ELISA. Result The results showed that the A sites and D sites of S gene and N genes were amplified, and their sizes were 498bp, 606bp and 1149bp, respectively. the nucleic acid vaccine expression vectors p-S(A-D)-His and p-N-His for A and D sites (fusion) and N gene (alone) were constructed. Bioinformatics software predicted that when TGEV infected host cells, N protein was mainly located in the nucleus and mitochondria, S(A-D) protein was mainly located in the cytoplasm and mitochondria, S(A-D) protein had 7 dominant B cell epitopes, and N protein had 8 dominant B cell epitopes. Recombinant vectors p-S(A-D)-His and p-N-His were successfully expressed in PK-15 cells, and S(A-D)-His and N-His were distributed in the nucleus and cytoplasm of PK-15 cells. After immunizing mice with vaccine vector, the immune effect was p-N-His ＞ p-S(A-D)-His + p-N-His ＞ p-S(A-D)-His from high to low. Conclusion In this paper, the DNA vaccine vectors of S and N genes of TGEV were constructed, and strong specific antibodies were generated after immunizing mice, which provided the basic material and basis for the development of nucleic acid vaccine of TGEV.
- TGEV,
- S gene、N gene,
- vector construction,
- immunogenicity

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References(22)

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