Given the possibility of cultural differences in the meaning and levels of gratitude among children, we evaluated the measurement invariance of the Gratitude Questionnaire–5 (GQ–5) and differences in latent means across adolescents from two distinct cultures, China and America. Data were obtained from 1,991 Chinese and 1,685 American adolescents. Confirmatory factor analysis and multigroup confirmatory factor analysis were performed to examine the factor structure and the measurement equivalence across Chinese and American adolescents. The Cronbach’s alpha and Item-total Correlations of the GQ–5 were also evaluated. Results of confirmatory factor analyses provided support for the expected one-factor structure. Also, a series of multi-group confirmatory factor analyses supported full configural invariance, full metric invariance, and partial scalar invariance between the two groups. Furthermore, the findings suggested that the GQ–5 is suitable for conducting mean level comparisons. The subsequent comparison of latent means revealed that the Chinese adolescents reported significantly lower gratitude than American adolescents.