{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T01:23:59Z","timestamp":1777512239854,"version":"3.51.4"},"reference-count":33,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2023,7,27]],"date-time":"2023-07-27T00:00:00Z","timestamp":1690416000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Jiangsu Agriculture Science and Technology Innovation Fund","award":["CX(21)3130"],"award-info":[{"award-number":["CX(21)3130"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>In this study, we applied bacterial artificial chromosome (BAC) technology with PRV\u0394TK\/gE\/gI as the base material to replace the first, central, and terminal segments of the US3 gene with codon-deoptimized fragments via two-step Red-mediated recombination in E. coli GS1783 cells. The three constructed BACs were co-transfected with gI and part of gE fragments carrying homologous sequences (gI+gE\u2019), respectively, in swine testicular cells. These three recombinant viruses with US3 codon de-optimization ((PRV\u0394TK&amp;gE-US3deop\u22121, PRV\u0394TK&amp;gE-US3deop\u22122, and PRV\u0394TK&amp;gE-US3deop\u22123) were obtained and purified. These three recombinant viruses exhibited similar growth kinetics to the parental AH02LA strain, stably retained the deletion of TK and gE gene fragments, and stably inherited the recoded US3. Mice were inoculated intraperitoneally with the three recombinant viruses or control virus PRV\u0394TK&amp;gEAH02 at a 107.0 TCID50 dose. Mice immunized with PRV\u0394TK&amp;gE-US3deop\u22121 did not develop clinical signs and had a decreased virus load and attenuated pathological changes in the lungs and brain compared to the control group. Moreover, immunized mice were challenged with 100 LD50 of the AH02LA strain, and PRV\u0394TK&amp;gE-US3deop\u22121 provided similar protection to that of the control virus PRV\u0394TK&amp;gEAH02. Finally, PRV\u0394TK&amp;gE-US3deop\u22121 was injected intramuscularly into 1-day-old PRV-negative piglets at a dose of 106.0 TCID50. Immunized piglets showed only slight temperature reactions and mild clinical signs. However, high levels of seroneutralizing antibody were produced at 14 and 21 days post-immunization. In addition, the immunization of PRV\u0394TK&amp;gE-US3deop\u22121 at a dose of 105.0 TCID50 provided complete clinical protection and prevented virus shedding in piglets challenged by 106.5 TCID50 of the PRV AH02LA variant at 1 week post immunization. Together, these findings suggest that PRV\u0394TK&amp;gE-US3deop\u22121 displays great potential as a vaccine candidate.<\/jats:p>","DOI":"10.3390\/vaccines11081288","type":"journal-article","created":{"date-parts":[[2023,7,28]],"date-time":"2023-07-28T01:51:47Z","timestamp":1690509107000},"page":"1288","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["A Novel Strategy of US3 Codon De-Optimization for Construction of an Attenuated Pseudorabies Virus against High Virulent Chinese Pseudorabies Virus Variant"],"prefix":"10.3390","volume":"11","author":[{"given":"Mengwei","family":"Xu","sequence":"first","affiliation":[{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Institute of Veterinary Immunology and Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"},{"name":"GuoTai (Taizhou) Center of Technology Innovation for Veterinary Biologicals, Taizhou 225300, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of the Ministry of Science and Technology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"},{"name":"College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yiwei","family":"Wang","sequence":"additional","affiliation":[{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Institute of Veterinary Immunology and Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"},{"name":"GuoTai (Taizhou) Center of Technology Innovation for Veterinary Biologicals, Taizhou 225300, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of the Ministry of Science and Technology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yamei","family":"Liu","sequence":"additional","affiliation":[{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Institute of Veterinary Immunology and Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"},{"name":"GuoTai (Taizhou) Center of Technology Innovation for Veterinary Biologicals, Taizhou 225300, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of the Ministry of Science and Technology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Saisai","family":"Chen","sequence":"additional","affiliation":[{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Institute of Veterinary Immunology and Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"},{"name":"GuoTai (Taizhou) Center of Technology Innovation for Veterinary Biologicals, Taizhou 225300, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of the Ministry of Science and Technology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Laixu","family":"Zhu","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ling","family":"Tong","sequence":"additional","affiliation":[{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Institute of Veterinary Immunology and Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"},{"name":"GuoTai (Taizhou) Center of Technology Innovation for Veterinary Biologicals, Taizhou 225300, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of the Ministry of Science and Technology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yating","family":"Zheng","sequence":"additional","affiliation":[{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Institute of Veterinary Immunology and Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"},{"name":"GuoTai (Taizhou) Center of Technology Innovation for Veterinary Biologicals, Taizhou 225300, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of the Ministry of Science and Technology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nikolaus","family":"Osterrieder","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Virologie, Freie Universit\u00e4t Berlin, 14163 Berlin, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chuanjian","family":"Zhang","sequence":"additional","affiliation":[{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Institute of Veterinary Immunology and Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"},{"name":"GuoTai (Taizhou) Center of Technology Innovation for Veterinary Biologicals, Taizhou 225300, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of the Ministry of Science and Technology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jichun","family":"Wang","sequence":"additional","affiliation":[{"name":"National Research Center of Engineering and Technology for Veterinary Biologicals, Institute of Veterinary Immunology and Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"},{"name":"GuoTai (Taizhou) Center of Technology Innovation for Veterinary Biologicals, Taizhou 225300, China"},{"name":"Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China"},{"name":"Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of the Ministry of Science and Technology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,7,27]]},"reference":[{"key":"ref_1","first-page":"99","article-title":"Aujeszky\u2019s disease (pseudorabies) virus: The virus and molecular pathogenesis\u2014State of the art, June 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