This paper aims at testing the influence of emission factors on travelers’ behavior of route choice. The generalized travel cost is defined as the linear weighted sum of emission factors, travel time, and travel time reliability. The relational model of exhaust volume and traffic volume is established using the BPR (Bureau of Public Road) function to calculate the cost of travel regarding emission. The BPR function is used to measure the road segment travel time, while the reliability is used to quantify the cost of travel time fluctuation. At last, the route choice model considering the generalized travel cost is established based on the game theory. The calculating and analyzing of results under a miniature road network show that the weight coefficient of travel cost influences the travelers’ behavior of route choice remarkably and the route choice model which takes emission into account can reduce the exhaust of road network effectively, approximately 11.4% in this case.