Vector Construction and Immunogenicity of <i>S</i> and <i>N</i> Gene DNA Vaccine 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. Vector Construction and Immunogenicity of S and N Gene DNA Vaccine for TGEV [J]. Fujian Journal of Agricultural Sciences，2024，39（9）：1−9

Citation:

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

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

Citation:

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

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Vector Construction and Immunogenicity of S and N Gene DNA Vaccine 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 DNA vaccine vector of S and N genes of porcine transmissible gastroenteritis virus (TGEV) was constructed with the vaccine immunogenicity detemined 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 498bp and 606bp, respectively, and the N, 1,149bp 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.
- TGEV,
- S and N gene,
- vector construction,
- immunogenicity

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References(22)

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