Abstract There have been some animal and human data suggesting that excitatory amino acid transporter (EAAT)-2, the major subtype of EAAT, is involved in human mental function and behavior. Recently, it has been shown that the −181 A/C polymorphism in the EAAT2 gene promoter affects plasma glutamate concentrations in humans. In the present study, the association of this genetic polymorphism with personality traits was examined in 575 Japanese healthy volunteers. Personality traits were assessed by the Temperament and Character Inventory, and the EAAT2 polymorphism was detected by a PCR–RFLP method. The scores of reward dependence were significantly ( p = 0.017) lower in the group with the A allele (A/A and A/C) than in that without this allele (C/C). When males and females were analyzed separately, the significant difference between the two genotype groups was observed in females ( p = 0.021) but not in males. The present study thus suggests that the −181 A/C polymorphism in the EAAT2 gene promoter affects the personality trait of reward dependence in healthy subjects.