[Objective] The function of the uncharacterized gene Afu4g13170 in Aspergillus fumigatus was ascertained. [Methods] Double-joint PCR method and one-step gene deletion technique were used to generate the A. fumigatus ΔAfu4g13170 mutant. [Results] Amino acid sequence alignment shows that the predicated Afu4g13170 protein has a similarity of 88.6% to Gib2 protein in Cryptococcus neoformans and Ani04163 protein in A. nidulans. The phenotypic analysis indicated that deletion of Afu4g13170 gene results in growth defect and incomplete development of phialides with elongated metulae, delayed and reduced sporulation, and less pigment production. Chromatographic analysis suggested that gliotoxin production is blocked partly by knocking out the Afu4g13170 gene. [Conclusion] Afu4g13170 gene may be used as one of target sites to control the virulence of A. fumigatus.