[Objectives]  To validate the expression of the Nosema ceranae γ-tubulin gene, clarify its molecular characteristics, and conduct a conserved motif and phylogenetic investigation of γ-tubulin proteins in N. ceranae and other species. [Methods]  The expression of γ-tubulin was verified by PCR. Its physical and chemical properties, signal peptide, phosphorylation site and secondary and tertiary structures, were predicted and analyzed using related software on the Expasy website. MEME software was used to predict conserved motifs in the γ-tubulin of N. ceranae and other species. Mega 11.0 software was used to perform multiple alignment of amino acid sequences of γ-tubulin from N. ceranae and other species, followed by construction of a phylogenetic tree based on the neighbor-joining method. [Results]  The expression of γ-tubulin was confirmed in N. ceranae spores. The molecular formula of the gene is C₂₁₈₆H₃₄₂₂N₅₈₂O₆₆₃S₂₀. It contains 433 amino acids and has a molecular weight of about 49.11 kD. Its lipid solubility coefficient is 86.65, and its hydrophilic coefficient is 5.75. The N. ceranae γ-tubulin gene contains 29 serine, 5 tyrosine and 14 threonine phosphorylation sites, and also includes a 191 α-helix, 66 extension, 17 β-turn and 159 random coil. The gene is found in the nucleus, cytoplasm and mitochondria. The same five conserved motifs were identified in the γ-tubulin gene of N. ceranae and those of another nine species. Phylogenetic analysis indicates that the γ-tubulin gene of N. ceranae, Nosema apis, Nosema bombycis, Encephalitozoon cuniculi, Encephalitozoon hellem and Vittaforma corneae cluster within a single large clade, whereas that of Rhizopus microsporus and Rhizopus azygosporus cluster in another large clade. The closest evolutionary distance was that between the γ-tubulin protein of N. ceranae and that of its sister species N. apis. [Conclusion]  Our results confirm the expression of γ-tubulin in N. ceranae spores, clarify the molecular characteristics of this gene, reveal a close genetic relationship between γ-tubulin in N. ceranae and that in N. apis, and show that γ-tubulin is highly conserved in N. ceranae, N. apis, N. bombycis, E. cuniculi, E. hellem and V. corneae.