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In this study, we isolated the FCV\u2010BJ616 strain, established a reverse\u2010genetics system, and investigated its pathogenic mechanisms, thereby providing a foundation for antibody\u2010based therapies and broad\u2010spectrum vaccine development. The virus was purified by three rounds of plaque cloning, and its morphology was examined by electron microscopy. VP1 expression was confirmed by immunofluorescence and Western blotting. Using integrated systems\u2010biology and reverse\u2010genetics approaches, an infectious clone of rFCV\u2010BJ616 was successfully assembled and rescued, exhibiting genetic stability comparable to that of the parental strain. In vivo infection experiments showed that rFCV\u2010BJ616 retained wild\u2010type virulence, causing persistent high fever, weight loss, and multiorgan pathology in infected cats. Proteomic analysis indicated that infection with FCV\u2010BJ616 or rFCV\u2010BJ616 markedly activated cytokine\u2010mediated inflammatory signaling pathways. Both FCV\u2010BJ616 and rFCV\u2010BJ616 significantly upregulated the expression of IL\u20108, S100A8\/A9, and TLR3, which are associated with acute inflammation and tissue damage. Furthermore, elevated IFN\u2010\u03b2 levels concomitant with STAT1 downregulation suggested a transient attenuation of antiviral signaling during early immune activation. These findings were corroborated by ELISA\u2010based validation of serum cytokine profiles. Collectively, this study provides new insights into the molecular pathogenesis and evolution of FCV\u2010BJ616 and establishes a robust reverse\u2010genetics platform for precise genome manipulation and future vaccine development.<\/jats:p>","DOI":"10.1002\/mbo3.70226","type":"journal-article","created":{"date-parts":[[2026,1,28]],"date-time":"2026-01-28T13:14:52Z","timestamp":1769606092000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Construction of Reverse Genetics System for Feline Calicivirus FCV\u2010BJ616 and Proteomic Analysis"],"prefix":"10.1002","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7833-5068","authenticated-orcid":false,"given":"Chunmei","family":"Xu","sequence":"first","affiliation":[{"name":"Key Laboratory of Animal Biosafety Risk Prevention and Control (North) &amp; Key Laboratory of Veterinary Biological Products and Chemical Drugs of MARA, Institute of Animal Science, 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