Altered expressions of mitochondria elongation factor Tu (EF-Tu) have been observed in certain types of cancers, including gastric cancer cell lines, but the impact of the alterations in gastric adenocarcinoma remains unclear. The purpose of this study was to investigate the expression of EF-Tu in gastric adenocarcinoma and to assess its clinical significance. A total of 104 paired resected gastric adenocarcinoma and corresponding normal specimens were collected in this study. EF-Tu expression was assessed by immunohistochemical staining. The correlation of EF-Tu expression and patients' clinicopathological parameters was statically evaluated and the prognostic significance of EF-Tu expression was assessed by univariate and multivariate analyses. Forty-nine out of 104 (47.1%) gastric adenocarcinoma specimens showed high expression of EF-Tu, while the remaining 55 specimens showed weak or negative expression of EF-Tu. In contrast, EF-Tu high expression was detected in 62.5% (65 of 104) normal tissues. Down-regulation of EF-Tu was associated with serosal invasion (P = 0.042) and node involvement (P = 0.005), and down-regulation of EF-Tu was correlated with poor overall survival (P = 0.020). In curative resection (R0) patients, there were also significant differences(P = 0.043). In the multivariate analysis, the EF-Tu expression remained a significant independent prognostic factor (P = 0.038). Our results indicate that EF-Tu is expressed in both gastric adenocarcinoma and corresponding normal tissues. Down-regulation of EF-Tu expression is associated with advanced disease stage and EF-Tu expression maybe served as an independent prognostic factor.