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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

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

  • Received Date: 2024-06-03
  • Rev Recd Date: 2024-08-23
  • Available Online: 2024-10-29
  •   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.
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  • [1]
    张羽欣, 王树茂, 段宏勇, 等. 猪传染性胃肠炎病毒TaqMan实时荧光定量PCR检测方法的建立与应用 [J]. 中国兽医科学, 2024, 54(4):479−484.

    ZHANG Y X, WANG S M, DUAN H Y, et al. Establishment and application of TaqMan real-time quantitative PCR for detection of porcine transmissible gastroenteritis virus [J]. Chinese Veterinary Science, 2024, 54(4): 479−484. (in Chinese)
    [2]
    JI Z Y, DONG H, JIAO R X, et al. The TGEV membrane protein interacts with HSC70 to direct virus internalization through clathrin-mediated endocytosis [J]. Journal of Virology, 2023, 97(4): e0012823. doi: 10.1128/jvi.00128-23
    [3]
    PU J N, CHEN D W, TIAN G, et al. All-trans retinoic acid attenuates transmissible gastroenteritis virus-induced inflammation in IPEC-J2 cells via suppressing the RLRs/NF-κB signaling pathway [J]. Frontiers in Immunology, 2022, 13: 734171. doi: 10.3389/fimmu.2022.734171
    [4]
    NIU Z, XU S S, ZHANG Y L, et al. Transmissible gastroenteritis virus nucleocapsid protein interacts with Na+/H+ exchanger 3 to reduce Na+/H+ exchanger activity and promote piglet diarrhea [J]. Journal of Virology, 2022, 96(22): e0147322. doi: 10.1128/jvi.01473-22
    [5]
    QIAN J T, LI M J, FENG Y F, et al. Genetic epidemiology of porcine transmissible gastroenteritis virus based on whole genome and S gene sequences[C]//2021 IEEE 9th International Conference on Bioinformatics and Computational Biology (ICBCB). May 25-27, 2021, Taiyuan, China. IEEE, 2021: 148-151.
    [6]
    郝振业. 猪传染性胃肠炎病毒反向遗传操作系统的构建及附属蛋白3(ORF3)的定位及功能研究[D]. 哈尔滨: 东北农业大学, 2023.

    HAO Z Y. Construction of reverse genetic operating system of porcine transmissible gastroenteritis virus and study on location and function of accessory protein 3(ORF3)[D]. Harbin: Northeast Agricultural University, 2023. (in Chinese)
    [7]
    王艳春. 猪传染性胃肠炎病毒S基因A位点杆状病毒表达及初步应用[D]. 扬州: 扬州大学, 2020.

    WANG Y C. Expression of baculovirus at site A of S gene of porcine infectious gastroenteritis virus and its preliminary application[D]. Yangzhou: Yangzhou University, 2020. (in Chinese)
    [8]
    张海燕. PEDV和TGEV的S蛋白融合抗原表位核酸疫苗的研究[D]. 武汉: 华中农业大学, 2019.

    ZHANG H Y. Study on S protein fusion epitope nucleic acid vaccine of PEDV and TGEV[D]. Wuhan: Huazhong Agricultural University, 2019. (in Chinese)
    [9]
    LI X L, LI P C, CAO L Y, et al. Porcine IL-12 plasmid as an adjuvant improves the cellular and humoral immune responses of DNA vaccine targeting transmissible gastroenteritis virus spike gene in a mouse model [J]. Journal of Veterinary Medical Science, 2019, 81(10): 1438−1444. doi: 10.1292/jvms.18-0682
    [10]
    师一鸣. TGEV单克隆抗体的制备及检测方法的建立[D]. 哈尔滨: 东北农业大学, 2019.

    SHI Y M. Preparation of TGEV monoclonal antibody and establishment of detection method[D]. Harbin: Northeast Agricultural University, 2019. (in Chinese)
    [11]
    李雅静, 宫强. 铜绿假单胞菌oprH基因DNA疫苗的构建与检测 [J]. 现代畜牧兽医, 2022(11):19−23. doi: 10.3969/j.issn.1672-9692.2022.11.lnxmsy202211006

    LI Y J, GONG Q. Construction and detection of Pseudomonas aeruginosa oprH gene DNA vaccine [J]. Modern Journal of Animal Husbandry and Veterinary Medicine, 2022(11): 19−23. (in Chinese) doi: 10.3969/j.issn.1672-9692.2022.11.lnxmsy202211006
    [12]
    杨鹏, 吴燕, 岳筠, 等. 绵羊肺炎支原体P113蛋白C末端基因真核表达载体的构建及其小鼠免疫应答 [J]. 中国兽医学报, 2022, 42(3):496−501, 521.

    YANG P, WU Y, YUE J, et al. Construction of eukaryotic expression vector for C terminal gene of Mycoplasma ovipneumoniae P113 protein and its immune response in mice [J]. Chinese Journal of Veterinary Science, 2022, 42(3): 496−501, 521. (in Chinese)
    [13]
    姚思, 杨洁琼, 杨雨欣, 等. 结核分枝杆菌ESAT6-Fc DNA疫苗的免疫效应评价 [J]. 中国生物制品学杂志, 2023, 36(8):897−901.

    YAO S, YANG J Q, YANG Y X, et al. Evaluation of immune effect of ESAT6-Fc DNA vaccine against Mycobacterium tuberculosis [J]. Chinese Journal of Biologicals, 2023, 36(8): 897−901. (in Chinese)
    [14]
    黄小波, 杨恒, 曹三杰, 等. 猪传染性胃肠炎病毒S-N融合双基因疫苗的构建及其免疫原性分析 [J]. 中国兽医科学, 2012, 42(8):848−853.

    HUANG X B, YANG H, CAO S J, et al. Construction and immunogenicity analysis of the S-N fusion gene vaccine against porcine transmissible gastroenteritis virus [J]. Chinese Veterinary Science, 2012, 42(8): 848−853. (in Chinese)
    [15]
    WANG G, LIANG R, LIU Z W, et al. The N-terminal domain of spike protein is not the enteric tropism determinant for transmissible gastroenteritis virus in piglets [J]. Viruses, 2019, 11(4): 313. doi: 10.3390/v11040313
    [16]
    韩郁茹, 石达, 张记宇, 等. 猪急性腹泻综合征冠状病毒RT-LAMP快速检测方法的建立与应用 [J]. 中国预防兽医学报, 2021, 43(1):35−39.

    HAN Y R, SHI D, ZHANG J Y, et al. Development and application of RT-LAMP method for rapid detection of SADS-CoV [J]. Chinese Journal of Preventive Veterinary Medicine, 2021, 43(1): 35−39. (in Chinese)
    [17]
    樊杰. 猪传染性胃肠炎病毒N蛋白纳米抗体的制备和基于纳米抗体竞争ELISA的建立[D]. 杨凌: 西北农林科技大学, 2021.

    FAN J. Preparation of nano-antibody against N protein of transmissible gastroenteritis virus and establishment of competitive ELISA based on nano-antibody[D]. Yangling: Northwest A & F University, 2021. (in Chinese)
    [18]
    ZHANG Y D, ZHANG X H, LIAO X D, et al. Construction of a bivalent DNA vaccine co-expressing S genes of transmissible gastroenteritis virus and porcine epidemic diarrhea virus delivered by attenuated Salmonella typhimurium [J]. Virus Genes, 2016, 52(3): 354−364. doi: 10.1007/s11262-016-1316-z
    [19]
    何雷, 董玲娟, 张彦明, 等. 猪传染性胃肠炎病毒ORF7蛋白在ST细胞中定位及其对病毒复制影响的研究 [J]. 中国预防兽医学报, 2020, 42(6):543−548.

    HE L, DONG L J, ZHANG Y M, et al. The subcellular location of transmissible gastroenteritis virus protein ORF7 and its effect on viral replication [J]. Chinese Journal of Preventive Veterinary Medicine, 2020, 42(6): 543−548. (in Chinese)
    [20]
    何雷, 贾艳艳, 郁川, 等. 稳定表达猪传染性胃肠炎病毒N蛋白的ST细胞株的构建及其亚细胞定位 [J]. 中国预防兽医学报, 2016, 38(2):101−104. doi: 10.3969/j.issn.1008-0589.2016.02.04

    HE L, JIA Y Y, YU C, et al. Establishment of stably-expressed transmissible gastroenteritis virus N protein cell line and its subcellular location [J]. Chinese Journal of Preventive Veterinary Medicine, 2016, 38(2): 101−104. (in Chinese) doi: 10.3969/j.issn.1008-0589.2016.02.04
    [21]
    韩涛涛, 黎露, 唐青海, 等. 不同佐剂对猪传染性胃肠炎病毒S蛋白和猪流行性腹泻病毒S蛋白免疫原性的影响 [J]. 中国农学通报, 2020, 36(30):143−150. doi: 10.11924/j.issn.1000-6850.casb20191000740

    HAN T T, LI L, TANG Q H, et al. Different adjuvants: Effects on S protein immunogenicity of porcine transmissible gastroenteritis virus and porcine epidemic diarrhea virus [J]. Chinese Agricultural Science Bulletin, 2020, 36(30): 143−150. (in Chinese) doi: 10.11924/j.issn.1000-6850.casb20191000740
    [22]
    伊立超. PEDV和TGEV受体结合区基因在昆虫杆状病毒系统的表达与免疫原性分析[D]. 延吉: 延边大学, 2022.

    YI L C. Expression and immunogenicity analysis of PEDV and TGEV receptor binding region genes in insect baculovirus system[D]. Yanji: Yanbian University, 2022. (in Chinese)
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