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<p>In this study, <i>VP1</i> genes from duck hepatitis A virus (DHAV) genotypes 1 (DHAV-1) and 3 (DHAV-3) were amplified through reverse transcription polymerase chain reaction (RT-PCR) by using viral RNA as the template. The amplified genes were cloned into the plasmid pNZ8149 through homologous recombination, yielding the recombinant plasmids pNZ8149-DHAV-1 VP1 and pNZ8149-DHAV-3 VP1. These plasmids were electroporated into competent <i>L. lactis</i> NZ3900 cells to generate the recombinant strains <i>L. lactis</i> pNZ8149-DHAV-1 VP1/NZ3900 and <i>L. lactis</i> pNZ8149-DHAV-3 VP1/NZ3900, respectively. Successful expression of VP1 in the recombinant <i>L. lactis</i> strains was confirmed through Western blot. One-day-old ducklings were orally immunized with the recombinant <i>L. lactis</i> strains (1 × 10<sup>11</sup>&#xa0;CFU/200&#xa0;μL) three times. Serum and intestinal mucosal scrapings were collected on days 5, 10, and 15 after immunization and on day 7 after viral challenge. Levels of specific antibodies and cytokines were measured in these samples. The results indicated that the recombinant <i>L. lactis</i> strains were successfully constructed and that they effectively expressed VP1. After oral immunization, antibody and cytokine levels in the serum and intestinal mucosa were significantly higher in immunized ducklings than in nonimmunized ducklings. These findings indicated that the recombinant <i>L. lactis</i> strains effectively stimulated systemic and mucosal immune responses, conferring significant immunoprotection against viral infection in ducklings.</p>

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Construction of recombinant Lactococcus lactis expressing VP1 from duck hepatitis a virus types 1 and 3 and evaluation of its immune effect

  • 期刊论文

  • 发表日期：

    2026年2月21日

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  • Yafei Qin,
  • Yaru Fan,
  • Meijuan Zhang,
  • Saisai Zhao,
  • Xinhong Man,
  • Dalin He,
  • Youxiang Diao,
  • Junxun Li,
  • Jingdong Hu,
  • Yi Tang

摘要

In this study, VP1 genes from duck hepatitis A virus (DHAV) genotypes 1 (DHAV-1) and 3 (DHAV-3) were amplified through reverse transcription polymerase chain reaction (RT-PCR) by using viral RNA as the template. The amplified genes were cloned into the plasmid pNZ8149 through homologous recombination, yielding the recombinant plasmids pNZ8149-DHAV-1 VP1 and pNZ8149-DHAV-3 VP1. These plasmids were electroporated into competent L. lactis NZ3900 cells to generate the recombinant strains L. lactis pNZ8149-DHAV-1 VP1/NZ3900 and L. lactis pNZ8149-DHAV-3 VP1/NZ3900, respectively. Successful expression of VP1 in the recombinant L. lactis strains was confirmed through Western blot. One-day-old ducklings were orally immunized with the recombinant L. lactis strains (1 × 1011 CFU/200 μL) three times. Serum and intestinal mucosal scrapings were collected on days 5, 10, and 15 after immunization and on day 7 after viral challenge. Levels of specific antibodies and cytokines were measured in these samples. The results indicated that the recombinant L. lactis strains were successfully constructed and that they effectively expressed VP1. After oral immunization, antibody and cytokine levels in the serum and intestinal mucosa were significantly higher in immunized ducklings than in nonimmunized ducklings. These findings indicated that the recombinant L. lactis strains effectively stimulated systemic and mucosal immune responses, conferring significant immunoprotection against viral infection in ducklings.