Trichophyton mentagrophytes is a common fungal pathogen that causes human and animal dermatophytosis. Previous studies have shown that zinc deficiency inhibits T. mentagrophytes growth, and the ZafA gene of T. mentagrophytes can code the functionally similar zinc finger transcriptional factor that can promote zinc ion absorption; however, the impact of ZafA on virulence and pathogenicity remains undetermined. To assess its gene function, the ZafA mutant, ZafA-hph, and the ZafA complemented strain, ZafA+bar, were constructed via Agrobacterium tumefaciens-mediated transformation. Polymerase chain reaction and Southern blot analyses were used to confirm the disruption. In vitro growth capacity and virulence analyses comparing ZafA-hph with wild-type T. mentagrophytes and ZafA+bar showed that ZafA-hph’s growth performance, reproduction ability, and zinc ion absorption capacity were significantly lower than the wild-type T. mentagrophytes and ZafA+bar. ZafA-hph also showed weak hair biodegradation ability and animal pathogenicity. Thus, the significant decrease in T. mentagrophytes’ growth ability and virulence was due to a lack of the zinc-responsive activity factor rather than the transformation process. This study confirmed that the T. mentagrophytes’ zinc-responsive activity factor plays important roles in the pathogen’s growth, reproduction, zinc ion absorption, and virulence. This factor is important and significant for effectively preventing and controlling T. mentagrophytes infections.